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Lamin B1

Lamin B1 is a type of nuclear lamina protein that plays a critical role in regulating gene expression, chromatin organization, and cellular differentiation.
It is essential for maintaining the structural integrity of the cell nucleus and is involved in various cellular processes, including DNA repair, transcription, and cell cycle regulation.
Lamin B1 has been extensively studied in the context of various diseases, such as premature aging disorders, neurodegenerative diseases, and cancer.
Researchers can optimize their Lamin B1 studies by utilizing the PubCompare.ai platform, which helps locate relevant protocols from literature, preprints, and patents, and provides AI-driven comparisons to identify the best protocols and products.
This can enhance the reproducibility and accuracy of Lamin B1 research, leading to advancements in our understanding of this important protein and its implications for human health.
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Most cited protocols related to «Lamin B1»

Quantifying Cell Cycle Protein Levels

Cited 186 times

Cells were grown on Histogrip (Invitrogen) coated glass coverslips and fixed using ice-cold 100% methanol (β-tubulin) or with 3.7% formaldehyde diluted in PBS with 0.5% Triton X-100 for 10 min (Mad2, pSerCdk, Lamin A/C, Plk1, cyclin B1, and securin). All cells were washed and then blocked (3% BSA, 0,1% Tween 20 in PBS) for 30 min. Cells were incubated with primary antibodies were incubated for 2 h at room temperature in blocking solution. DNA was stained with DAPI. For Lamin A/C staining a Leica DM6000 SP8 confocal with a 63× lens was used. All other images were captured using Leica DM5500 microscope coupled with a Coolsnap HQ2 camera, using a Leica 100× or 40× APO 1.4 lens, powered by Leica LAS AF v3 software. To quantify pSer-CDK, cyclin B and secruin levels in cells, a single in-focus plane was acquired. Using ImageJ (v1.48, NIH), an outline was drawn around each cell and circularity, area, mean fluorescence measured, along with several adjacent background readings. The total corrected cellular fluorescence (TCCF) = integrated density – (area of selected cell × mean fluorescence of background readings), was calculated. This TCCF was then equalized against the mean TCCF of neighboring interphase cells in the same field of view, with results presented as fold increase over interphase levels. Box plots and statistical analysis (2-sided unpaired Student t tests) were performed using GraphPad Prism 5. For all other images, 0.3 µm z-sections were taken, de-convolved, and displayed as 2D maximum projections using ImageJ. False coloring and overlays were performed using Adobe Photoshop CS5 software.

McCloy R.A., Rogers S., Caldon C.E., Lorca T., Castro A, & Burgess A. (2014). Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events. Cell Cycle, 13(9), 1400-1412.

Publication 2014

Antibodies Cells Cold Temperature Cyclin B Cyclin B1 DAPI Fluorescence Formaldehyde Interphase Lens, Crystalline LMNA protein, human Methanol Microscopy PLK1 protein, human prisma PTTG1 protein, human Student Triton X-100 Tubulin Tween 20

Constructing mNeonGreen Fusion Protein Vectors

Cited 67 times

mNeonGreen fluorescent protein expression vectors were constructed using C1 and N1 (Clontech-style) cloning vectors. The mNeonGreen cDNA was amplified with a 5′ primer encoding an AgeI site and a 3′ primer encoding either a BspEI (C1) or NotI (N1) site for generating cloning vectors to create C-terminal and N-terminal fusions (with regards to the FP), respectively. Purified and digested PCR products were ligated into similarly digested EGFP-C1 and EGFP-N1 cloning vector backbones. To obtain targeting fusion vectors, the appropriate cloning vector and a previously assembled EGFP or mEmerald fusion vector were digested, either sequentially or doubly, with the appropriate enzymes and ligated together after gel purification.
Thus, to prepare mNeonGreen C-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: annexin A4 (12), NheI and BspEI (Alen Piljic, EMBL, Heidelberg, Germany; NM_001153.3); β-actin (7), NheI and BglII (human β-actin cDNA source: Clontech; NM_001101.3); β-catenin (20), XhoI and BamHI (mouse β-catenin cDNA source: Origene, Rockville, MD; NM_001165902.1); 20 amino acid farnesylation signal from c-Ha-Ras (CAAX; 5), AgeI and BspEI (c-Ha-Ras cDNA source: Clontech; NM_001130442.1); CAF1 (10), AgeI and BspEI (mouse chromatin assembly factor cDNA source: Akash Gunjan, Florida State University; NM_013733.3); caveolin 1 (10), NheI and BglII (human caveolin 1 cDNA source: Origene; NM_001753); endosomes (14), NheI and BspEI (endosomes cDNA source: Clontech; NM_004040.2); fascin (10), BspEI and BamHI (human fascin cDNA source: Origene; NM_003088.2); fibrillarin (7), AgeI and BspEI (fibrillarin cDNA source: Evrogen, Moscow, Russia; NM_001436.3); filamin A (14), BspEI and HindIII (human filamin cDNA source: David Calderwood, Yale University; NM_001456.3); human lysosomal membrane glycoprotein 1 (20), BamHI and NotI (LAMP1; George Patterson, NIH, Bethesda MD, U.S.A.; NM_012857.1); human light chain clathrin (15), NheI and BglII (human clathrin light chain cDNA source: George Patterson, NIH; NM_001834.2); human myotilin, AgeI and BspEI (MYOT; Origene; NM_006790.1); PCNA (19), AgeI and BspEI (proliferating cell nuclear antigen cDNA source: David Gilbert, FSU; NM_002592.2); plastin (10), BspEI and XhoI (human plastin 1 (fimbrin) cDNA source: Origene; NM_002670.1); canine Rab4a, BglII and BamHI (Rab4a cDNA source: Viki Allen, U. Manchester, UK; NM_004578.2); LC3B (7), AgeI and BspEI (rat LC3B cDNA source: Jenny M. Tam, Harvard University; U05784.1); talin (22) AgeI and BspEI (mouse talin 1 cDNA source: Clare Waterman, NIH; NM_011602.5); α-tubulin (18), NheI and BglII (human α-tubulin cDNA source: Clontech; NM_006082).
To prepare mNeonGreen N-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: human non-muscle α-actinin, EcoRI and NotI (cDNA source, Tom Keller, Florida State University (FSU), Tallahassee, FL, U.S.A.; NM_001130005.1); human calnexin, AgeI and NotI (Origene; NM_001746.3); c-src (7), BamHI and EcoRI (chicken c-src cDNA source: Marilyn Resh, Sloan-Kettering, New York; XM_001232484.1); connexin-43 (7), BamHI and NotI (rat C×43 cDNA source: Matthias Falk, Lehigh U; NM_001004099.1); EB3 (7), BglII and BamHI (EB3 cDNA source: Lynne Cassimeris, Lehigh University; NM_012326.2); human keratin 18, EcoRI and NotI (Open Biosystems; NM_199187.1); lamin B1 (10), EcoRI and BamHI (human lamin B1 cDNA source: George Patterson, NIH; NM_005573.2); Lifeact (7), BamHI and NotI (Lifeact cDNA source: IDT); mouse mannosidase 2 (112 N-terminal amino acids, MANNII; 10), NheI and BamHI (cDNA source: Jennifer Lippincott-Schwartz, NIH; NM_008549.2); myosin IIA (14) NheI and BglII (mouse myosin IIA cDNA source: Origene; NM_022410.2); human nucleoporin 50kDa, BamHI and NotI (NUP50 cDNA source: Origene; NM_007172.2); human pyruvate dehydrogenase, AgeI and NotI (human PDHA1 cDNA source: Origene; NM_000284); human peroxisomal membrane protein, NotI and AgeI (PMP cDNA source: Origene; NM_018663.1); human MAP Tau (10), AgeI and NotI (MAP Tau cDNA source: Origene; NM_016841); human TfR (20), BamHI and NotI (transferrin receptor cDNA source: George Patterson, NIH; NM_NM_003234); human TPX2 (10), AgeI and NotI (TPX2 cDNA source: Patricia Wadsworth, University of Massachusetts, Amherst; NM_012112.4); mouse VASP (10), NheI and BamHI (cDNA source: Clare Waterman, NIH; NM_009499); vascular epithelial cadherin (10), AgeI and NotI (human VE cadherin cDNA source: Origene; NM_001795.3), vimentin (7), BamHI and NotI (human vimentin cDNA source: Robert Goldman, Northwestern University; NM_003380.3), zyxin (6), BamHI and NotI (human zyxin cDNA source: Origene; NM_003461). All DNA for transfection was prepared using the Plasmid Maxi kit (QIAGEN). To ensure proper localization, mNeonGreen fusion proteins were characterized by transfection in HeLa (S3 or CCL2 line) or MDCK cells (ATCC) using Effectene (QIAGEN) and ∼1 μg vector. Transfected cells were grown on coverslips in DMEM/F12, fixed after 48 hours, and mounted with Gelvatol. Epifluorescence images were captured with a Nikon 80i microscope using widefield illumination and a Chroma FITC filter set to confirm proper localization.

Shaner N.C., Lambert G.G., Chammas A., Ni Y., Cranfill P.J., Baird M.A., Sell B.R., Allen J.R., Day R.N., Israelsson M., Davidson M.W, & Wang J. (2013). A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nature methods, 10(5), 407-409.

Publication 2013

Construction and Characterization of mTagBFP2 Fusion Proteins

Cited 22 times

mTagBFP2 fluorescent protein expression vectors were constructed using -C1 and -N1 (Clontech-style) cloning vectors. The mTagBFP2 cDNA was amplified with a 5′ primer encoding an AgeI site and a 3′ primer encoding either a BspEI (-C1) or NotI (-N1) site for C-terminal and N-terminal fusions (with regards to the FP), respectively. Purified and digested PCR products were ligated into similarly digested pEGFP-C1 and pEGFP-N1 cloning vector backbones. To generate targeting fusion vectors, the appropriate cloning vector and a previously assembled EGFP fusion vector were digested, either sequentially or doubly, with the appropriate enzymes and ligated together after gel purification.
Thus, to prepare mTagBFP2 C-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: human lamin B1 (10), NheI and BglII (lamin B1 cDNA source: George Patterson, NIH; NM_005573.2); 20 amino acid farnesylation signal from c-Ha-Ras (CAAX; 5), AgeI and BspEI (c-Ha-Ras cDNA source: Clontech, Mountain View, CA; NM_001130442.1); endoplasmic reticulum (5), AgeI and BspEI (calreticulin cDNA source: George Patterson, NIH; NM_004343.3); fibrillarin (7), AgeI and BspEI (fibrillarin cDNA source: Evrogen, Moscow, Russia; NM_001436.3); human light chain clathrin (15), NheI and BglII (clathrin cDNA source: George Patterson, NIH; NM_001834.2); β-actin (7), NheI and BglII (human β-actin cDNA source: Clontech, Mountain View, CA; NM_001101.3); caveolin 1 (10), NheI and BglII (human caveolin 1 cDNA source: Origene, Rockville, MD; NM_001753); vinculin (22) AgeI and EcoRI (human vinculin source: Clare Waterman, NIH; NM_003373.3); CAF1 (10), AgeI and BspEI (mouse chromatin assembly factor cDNA source: Akash Gunjan, FSU; NM_013733.3) Rab5a (7), NheI and BglII (canine Rab5a cDNA source: Vicki Allen, University of Manchester; NM_001003317.1); α-tubulin (18), NheI and BglII (human α-tubulin cDNA source: Clontech, Mountain View, CA; NM_006082); myosin IIA (18) NheI and BglII (human myosin heavy chain IIA cDNA source: DNA2.0, Menlo Park, CA; AJ312390.1); PCNA (19), AgeI and BspEI (proliferating cell nuclear antigen cDNA source: David Gilbert, FSU; NM_002592.2).
To prepare mTagBFP2 N-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: β-2 connexin-26 (7), BamHI and NotI (rat Cx26 cDNA source: Matthias Falk, Lehigh U; NM_001004099.1); TfR (20), BamHI and NotI (transferrin receptor cDNA source: George Patterson, NIH; NM_NM_003234); Golgi complex (7), BamHI and NotI (human β-galactosamide α-2,6-sialyltransferase 1cDNA source: Jennifer Lippincott-Schwartz, NIH; NM_173216.2); zyxin (6), BamHI and NotI (human zyxin cDNA source: Origene, Rockville, MD; NM_003461); vascular epithelial cadherin (10), AgeI and NotI (human VE cadherin cDNA source: Origene, Rockville, MD; NM_001795.3); mitochondria (7), BamHI and NotI (human mitochondrial targeting sequence, cytochrome c oxidase cDNA source: Clontech, Mountain View, CA; NM_004074.2); centromere protein B (22), BamHI and NotI (human CENPB cDNA source: Alexey Khodjakov, Wadsworth Center, Albany, NY; NM_001810.5); α-actinin (19), BamHI and EcoRI (human α-actinin cDNA source: Tom Keller, Florida State University, Tallahassee; NM_001130005.1); c-src sarcoma (7), BamHI and EcoRI (chicken c-src cDNA source: Marilyn Resh, Sloan-Kettering, New York; XM_001232484.1); Lifeact (7), BamHI and NotI (Lifeact cDNA source: IDT, Coralville, IA); vimentin (7), BamHI and NotI (human vimentin cDNA source: Robert Goldman, Northwestern University; NM_003380.3).
All DNA for transfection was prepared using the Plasmid Maxi kit (QIAGEN, Valencia, CA). To ensure proper localization, mTagBFP2 fusion proteins were characterized by transfection in HeLa cells (CCL2 line; ATCC, Manassas, VA) using Effectene (QIAGEN) and 1 µg vector. Transfected cells were grown on coverslips in DMEM/F12, fixed after 48 hours, and mounted with Gelvatol. Epifluorescence images (Figure 4) were taken with a Nikon 80i microscope using widefield illumination and an Omega QMax Blue filter set to confirm proper localization.

Subach O.M., Cranfill P.J., Davidson M.W, & Verkhusha V.V. (2011). An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore. PLoS ONE, 6(12), e28674.

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Publication 2011

Chromatin Immunoprecipitation Protocol

Cited 18 times

Cells in 10-cm² dishes were fixed in 1% formaldehyde for 10 min, and fixation was quenched with addition of glycine to 125 mM for an additional 5 min. Cells were harvested by scraping from plates and washed twice in 1× PBS before storage at −80°C. ChIP was performed as described in the Young laboratory protocol (Lee et al. 2006b (link)), except that extracts were sonicated twice for 9 min each round (30 sec of sonication with intermediate incubation of 30 sec per round) using a Bioruptor (Diagenode). All ChIPs were performed using 500 μg of extract and 2 μg of antibody per sample. Thirty microliters of Protein G Dynabeads (Invitrogen, 100.02D) was used per ChIP. Controls with IgG and no antibody controls were routinely performed, and all antibodies were tested by titration to be functioning within the linear range of the protocol. Following elution, ChIP DNA was analyzed by standard qPCR methods on a 7900HT Fast-Real-Time PCR (ABI). Primer sequences are available on request. For sequencing, 10 ng of ChIP DNA was used to make sequencing libraries using standard Illumina library single-end construction procedures. Sequencing was performed on either Illumina GAIIx (36 base pairs [bp], single-end reads) or Hi-Seq (100 bp, paired-end or single-end reads) platforms.

Shah P.P., Donahue G., Otte G.L., Capell B.C., Nelson D.M., Cao K., Aggarwala V., Cruickshanks H.A., Rai T.S., McBryan T., Gregory B.D., Adams P.D, & Berger S.L. (2013). Lamin B1 depletion in senescent cells triggers large-scale changes in gene expression and the chromatin landscape. Genes & Development, 27(16), 1787-1799.

Publication 2013

Antibodies Cells DNA Chips DNA Library Formaldehyde G-substrate Glycine Hyperostosis, Diffuse Idiopathic Skeletal Immunoglobulins Oligonucleotide Primers Real-Time Polymerase Chain Reaction Titrimetry

Immunofluorescence Imaging of HeLa Cells

Cited 16 times

HeLa cells were grown on glass coverslips and treated as detailed in the figure legends. Cells were fixed in 2% paraformaldehyde/PHEM solution containing 0.5% Triton X-100 for 15 min. Coverslips were washed in PBST, blocked in 5%BSA/PBS, and incubated overnight with primary antibodies. Samples were then incubated with secondary antibodies for 2–3 h, stained with DNA dye, DAPI, and mounted using Vectashield (Vector Laboratories, Burlingame, CA). For data displayed in Figure 3 and Supplemental Figures 2 and 5, the following antibodies were used: mouse MPM2 (Dako Corp, Oregon City, OR), rabbit pS-Cdk (Cell Signaling) or mouse IgM pNucleolin (a gift from P. Davies). Each sample was coincubated with an antibody against the Lamin B1, either of mouse or of rabbit origin (both from Abcam). Secondary goat anti–rabbit and goat anti–mouse or anti–mouse IgM antibodies were conjugated to Cy3 and FITC (Jackson ImmunoResearch). DNA was stained with DAPI. The images were acquired using Zeiss Axiovert 200M wide-field fluorescence microscope (40× oil immersion objective) equipped with a Hamamatsu ORCA-ERG digital camera and processed with MetaMorph (Molecular Devices).
For data displayed in Figure 4, cells were labeled with rat antibody against tyrosinated alpha-tubulin (clone YL1/2; Abcam) followed by a secondary goat anti–rat antibody conjugated to Cy3. Subsequently, cells were labeled with mouse anti–pS10 Histone H3 antibody conjugated to Alexa Fluor 647 (Cell Signaling). DNA was stained with Vybrant DyeCycle Green (Molecular Probes). For data displayed in Supplemental Figure 3, cells were first labeled with primary mouse antibody against nucleolin (Abcam) and secondary goat anti–mouse antibody conjugated to Cy5. Subsequently, cells were labeled with phospho-Nucleolin mouse IgM antibody and the secondary antibody against mouse IgM conjugated to Cy3. DNA was stained with Vybrant DyeCycle Green. Images from these experiments were collected using a 63× PlanApochromat oil immersion objective on a Zeiss AxioObserver equipped with a high-speed Yokogawa CSU 22 spinning disk confocal imaging system and a Hamamatsu ORCA-ERG digital camera. Images were collected and processed with SlideBook software (Intelligent Imaging Innovations, Denver, CO).

Potapova T.A., Sivakumar S., Flynn J.N., Li R, & Gorbsky G.J. (2011). Mitotic progression becomes irreversible in prometaphase and collapses when Wee1 and Cdc25 are inhibited. Molecular Biology of the Cell, 22(8), 1191-1206.

Publication 2011

Alexa Fluor 647 alpha-Tubulin anti-IgM Antibodies Antibodies, Anti-Idiotypic Cells Clone Cells Cloning Vectors DAPI Fluorescein-5-isothiocyanate Goat HeLa Cells Histone H3 Immunoglobulin M Immunoglobulins Innovativeness lamin B1 Medical Devices Microscopy, Fluorescence Molecular Probes Mus nucleolin Orcinus orca paraform Rabbits Submersion Triton X-100

Most recents protocols related to «Lamin B1»

Lamin-B1 Immunofluorescence Microscopy

Lamin-B1 was detected by immunofluorescence microscopy. Cells were fixed with methanol/acetone at ratio 3/7 and stained overnight at 4°C with anti-Lamin-B1 rabbit mAb (Abcam, ab133741) diluted 1:100 in PBS with 5% BSA, 0.3 M glycine, 0.1% Triton X-100. After washing in PBS, AF594-cojugated goat anti-rabbit IgG secondary antibody (Invitrogen, A-11012) was applied for 1 h at room temperature. Cover slip was mounted using SlowFade™ gold antifade mountant with DAPI (Invitrogen, S36938). Images were acquired by conventional epifluorescence microscopy using an Olympus BX51 microscope equipped with a ProgRes® MF cool monochrome camera (Jenoptik) and processed with I.A.S software ver. 009 (Delta Sistemi) for merging and pseudo-coloring adjustment.

Favaretto G., Rossi M.N., Cuollo L., Laffranchi M., Cervelli M., Soriani A., Sozzani S., Santoni A, & Antonangeli F. (2024). Neutrophil-activating secretome characterizes palbociclib-induced senescence of breast cancer cells. Cancer Immunology, Immunotherapy : CII, 73(6), 113.

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Publication 2024

Immunofluorescent Imaging of Mouse Thymus

Not available on PMC !

Microscope slides containing mouse thymus sections were xed with ice-cold acetone and stained with recombinant anti-mouse lamin B1 antibody (clone EPR22165-121; Abcam) consisting of 10% HiFCS and 0.2% triton-X 100 in PBS overnight followed by staining with 6.6 µg/ml anti-rabbit IgG conjugated to Alexa Fluor ® 555 (clone H + L, F(ab') 2 ; Cell Signalling Technology). Post wash, tissues were stained with 1 µg/ml DAPI solution (Thermo Fischer ™ ). Images were acquired using an Olympus IX71 inverted uorescence microscope at 10X and 40X magni cation and imaging analysis was performed using Image J software.

Conway J., DeJong E.N., White A.J., Dugan B., Rees N.P., Parnell S.M., Lamberte L.E., Sharma‐Oates A., Sullivan J., Mauro C., Schaik W.v., Anderson G., Bowdish D.M.E., & Duggal N.A. (2024). Age-related loss of intestinal barrier integrity plays an integral role in Thymic involution and T cell ageing.

Publication 2024

HPV infection: Immunostaining and imaging protocol

For HPV infection studies, cells were fixed with 4% paraformaldehyde (freshly made) for 25 min followed by 3 × wash step with PBS (Life technologies, 14190-169). After permeabilization in 0.3% Triton X-100 (Sigma, X100-500 ml), cells were washed again 3 × with PBS before staining with DAPI (1 µg/ml) for 15 min. Plates were maintained in PBS-NaN₃ at 4 °C pending microscopic imaging. For IF, cells were blocked in 50% FBS after the permeabilization for 30 min and primary antibodies were added for one h. Following primary antibodies were used: mouse anti-lamin A/C (Santa Cruz Biotechnology, sc-376248, 1/100), rabbit anti-lamin B1 (Abcam, ab16048, 1/250), rabbit anti-lamin B2 (Abcam ab151735, 1/500), rabbit anti-PML (Santa Cruz, sc5621, 1/250), mouse anti-H3K9me2,3 (Cell Signaling Technology, #5327, 1/100), rabbit anti-H3K9ac (Cell Signaling Technology, #9649, 1/400), mouse anti-IRF3 (Abcam, ab68481, 1/100) and rabbit anti-p65 (Abcam, ab7970, 5 µg/ml). After 3 × 5 min wash step with PBS, the secondary antibodies; donkey anti-mouse CY3 (Jackson, 715-165-151, 1/600) and donkey anti-rabbit CY5 (Jackson, 711-175-152, 1/600), were added for 30 min. Afterwards cells were washed again 3 × 5 min with PBS and stained with DAPI similar as above. For transcription factor localization experiments additional HCS CellMask™ staining (Life Technologies, H32721, 1/5000) was applied, to allow distinguishing nuclear from cellular signal. To visualize EdU-labeled pseudogenomes, we made use of the Click-iT EdU Alexa Fluor™ 555 imaging kit (Invitrogen, C10338). Cells were incubated for 30 min at room temperature with the Click-iT reagent after blocking. Anonymized archival paraffin embedded human cervix samples, were microtome sectioned onto SuperFrost slides, deparaffinized (xylene) and rehydrated. After antigen retrieval (citrate buffer), they were subjected to an immunostaining for lamin A/C, lamin B1 and/or BAF (Abcam, ab129184, 1/500), counterstained with DAPI and mounted with Citifluor.

Molenberghs F., Verschuuren M., Vandeweyer L., Peeters S., Bogers J.J., Novo C.P., Vanden Berghe W., De Reu H., Cools N., Schelhaas M, & De Vos W.H. (2024). Lamin B1 curtails early human papillomavirus infection by safeguarding nuclear compartmentalization and autophagic capacity. Cellular and Molecular Life Sciences: CMLS, 81(1), 141.

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Publication 2024

Western Blot Protocol for Protein Analysis

Cells were grown in 6-well plates (Greiner Bio-One, 657102) or 12-well plates (Thermo Scientific, 150628) and lysed using M-PER® Mammalian Protein Extraction Reagent (Thermo scientific, 78503). Protein concentration was measured with the Pierce™ BCA Protein Assay Kit (Thermo scientific, 23227). Cell lysates were mixed with 25% NuPage LDS sample buffer (ThermoFisher, NP0007) and 5% dithiothreitol (DDT, ThermoFisher NP0009) and heated for 10 min on 70 °C. Samples (5 µg) were loaded onto NuPAGE™ Novex 4–12% Bis–Tris Protein Gels (ThermoFisher, NP0322PK2), with MOPS running buffer (Thermo Scientific, J00047). PageRuler P-prestained Protein Ladder was used as marker (ThermoFisher, PI26616). Next, proteins were transferred to BioTrace PVDF membranes (Pall Corporation, 66542) using a transfer mixture of NuPAGE transfer buffer (ThermoFisher), NuPAGE antioxidant (ThermoFisher) and methanol. Afterwards the membranes were blocked in blocking buffer (5% ECL (Sigma GERPN418) in Tris Buffered Saline with 0.2% Tween 20 (TBST)), and subsequently incubated with primary antibodies, diluted in blocking buffer. The following primary antibodies were used: mouse anti-lamin A/C (Santa Cruz Biotechnology, sc-376248, 1/100), rabbit anti-lamin B1 (Abcam, ab16048, 1/1000), rabbit anti-lamin B2 (Abcam ab151735, 1/1000), mouse anti-H3K9me2,3 (Cell Signaling Technology, #5327, 1/1000), rabbit anti-H3K9ac (Cell Signaling Technology, #9649, 1/1000) and anti-cGAS (Cell Signaling Technology, 15102S, 1/1000). Rabbit anti-Nucleolin (Novus Biologicals, NB600-241, 1/4000) and anti-GAPDH (GeneTex, GT239, 1/10000) were used as a reference protein. Horse radish peroxidase (HRP)-conjugated goat anti-mouse (Sigma-Aldrich A4416, 1/5000) and HRP-conjugated goat anti-rabbit (Sigma-Aldrich A6154, 1/5000) were used as secondary antibodies. Proteins were detected by chemiluminescence with Immobilon western chemiluminescent HRP substrate (Millipore, WBKLS0100) using a western blot Imager (Bio-Rad, ChemiDocTM XRS +). Quantification was done with Fiji image processing freeware [29 (link)] by measuring the intensity of each band in a rectangular selection of fixed size and the intensity of each marker band was expressed relative to that of the corresponding reference protein in the same lane.

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Publication 2024

Immunofluorescence Analysis of Lamin B1 and Fibronectin

Tissues (5 μm) were de-paraffinized in xylene and rehydrated with ethanol before antigen retrieval. After antigen retrieval in EDTA solution, the sections were washed with PBS for 15 min and treated with blocking buffer for 30 min at room temperature. For Lamin B1 staining, sections were incubated with anti-rabbit Lamin B1 antibody (1:200; AF5161-50; Affinity Biosciences; Melbourne, Australia) overnight at 4°C, and the fluorescently labeled secondary antibody (1:1000; coralite594, SA00013-4; Proteintech; Wuhan, Hubei, China) for 1 h at room temperature. For Lamin B1 and Fibronectin staining in cells, cells were seeded on glass slides and the slides were fixed in 4% formaldehyde in PBS for 15 min, washed three times in PBS, and then permeabilized with 0.1% TritonX-100 in PBS for 10 min. Slides were then blocked with 3% BSA in PBS for 1 h at room temperature followed by incubation with the primary antibody (Lamin B1, 1:200; Fibronectin, 1:200, 15613–1-AP, Proteintech) at 4°C overnight and the secondary antibody for 1 h at room temperature. Nuclei were stained with DAPI (HNFD-02; HelixGen). Images were taken with Olympus BX53 microscope.

Chen J., Li G., He X., Chen X., Chen Z., Liu D., Guo S., Huang T., Lin Y., Lan P., Lian L, & He X. (2024). ELMO1 ameliorates intestinal epithelial cellular senescence via SIRT1/p65 signaling in inflammatory bowel disease-related fibrosis. Gastroenterology Report, 12, goae045.

Publication 2024

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Ab16048 by Abcam

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Ab16048 is a monoclonal antibody that recognizes the protein Actin. This antibody can be used to detect and quantify Actin in various sample types.

Lamin b1 by Abcam

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Lamin B1 is a structural protein that is a key component of the nuclear lamina, a protein meshwork that provides mechanical support and organization to the cell nucleus. It plays an essential role in maintaining nuclear structure and integrity.

Lamin b1 by Cell Signaling Technology

Sourced in United States, United Kingdom

Lamin B1 is a nuclear envelope protein that is a structural component of the nuclear lamina. It plays a key role in maintaining the integrity and organization of the nucleus.

Lamin b1 by Santa Cruz Biotechnology

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Lamin B1 is a structural protein that is a component of the nuclear lamina, a protein network that provides mechanical support and organization to the cell nucleus. Lamin B1 is involved in the maintenance of nuclear structure and function.

Pvdf membrane by Merck Group

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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.

Ne per nuclear and cytoplasmic extraction reagent by Thermo Fisher Scientific

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The NE-PER Nuclear and Cytoplasmic Extraction Reagents are a set of buffers designed to facilitate the isolation of nuclear and cytoplasmic protein fractions from eukaryotic cells. The reagents enable the separation of these cellular compartments, allowing for further analysis or study of the extracted proteins.

Anti lamin b1 by Abcam

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Anti-Lamin B1 is a laboratory reagent used to detect the presence and localization of Lamin B1 protein, a structural protein found in the cell nucleus. It is commonly used in immunohistochemistry, immunocytochemistry, and Western blotting applications.

Lamin b1 by Proteintech

Sourced in United States, China, United Kingdom

Lamin B1 is a protein that is a component of the nuclear lamina, a meshwork of proteins found underneath the inner nuclear membrane in eukaryotic cells. Lamin B1 is involved in the structural organization of the nucleus and in regulating gene expression.

β actin by Merck Group

Sourced in United States, United Kingdom, Germany, China, Canada, Japan, Macao, Italy, Sao Tome and Principe, Israel, Spain, Denmark, France, Finland, Australia, Morocco, Ireland, Czechia, Sweden, Uruguay, Switzerland, Netherlands, Senegal

β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.

β actin by Cell Signaling Technology

Sourced in United States, United Kingdom, China, Germany, Japan, Canada, Morocco, Sweden, Netherlands, Switzerland, Italy, Belgium, Australia, France, India, Ireland

β-actin is a cytoskeletal protein that is ubiquitously expressed in eukaryotic cells. It is an important component of the microfilament system and is involved in various cellular processes such as cell motility, structure, and integrity.

What is Lamin B1 and what is its importance in cellular processes?

Lamin B1 is a critical nuclear lamina protein that plays a key role in regulating gene expression, chromatin organization, and cellular differentiation. It is essential for maintaining the structural integrity of the cell nucleus and is involved in various cellular processes, including DNA repair, transcription, and cell cycle regulation. Lamin B1 has been extensively studied in the context of diseases such as premature aging disorders, neurodegenerative diseases, and cancer, making it an important target for research and understanding human health.

How can PubCompare.ai help optimize Lamin B1 research?

PubCompare.ai can help researchers optimize their Lamin B1 studies in several ways. First, it allows you to screen protocol literature more efficently, helping you locate relevant protocols from literature, preprints, and patents. Second, the platform's AI-driven analysis can help you leverage critical insights, enabling you to identify the most effective protocols related to Lamin B1 for your specific research goals. This can enhance the reproducibility and accuracy of your Lamin B1 research, leading to advancements in our understanding of this important protein and its implications for human helath.

What are some common challenges in working with Lamin B1?

One of the key challenges in working with Lamin B1 is ensuring reproducibility and accuracy in your research. Lamin B1 is involved in complex cellular processes, and slight variations in experimental protocols can significantly impact your findings. Additionaly, there are different isoforms and variants of Lamin B1 that may behave differently in various cellular contexts. Leveraging a tool like PubCompare.ai can help you navigate these challenges by identifying the most effective protocols and products for your specific Lamin B1 research needs.

How can PubCompare.ai help researchers compare Lamin B1 protocols and products?

PubCompare.ai's AI-driven comparrison tools can be invaluable for Lamin B1 researchers. The platform allows you to compare protocols and products side-by-side, highlighting key differences in effectiveness, reproducibility, and other critical factors. This empowers you to make informed decisions about the best approaches to use in your Lamin B1 studies, enhancing the quality and impact of your research. Whether you're looking to optimize existing protocols or explore new techniques, PubCompare.ai can be a powerful ally in your Lamin B1 research journey.

What are some specific applications of Lamin B1 research?

Lamin B1 research has a wide range of applications, particularly in the context of human health and disease. Studeis of Lamin B1 have provided insights into premature aging disorders, neurodegenerative diseases, and cancer, as this protein plays a critical role in maintaining nuclear integrity and regulating cellular processes. Advancements in Lamin B1 research can lead to new therapeutic targets and improved understanding of the molecular mechanisms underlying these debilitating conditions. By leveraging tools like PubCompare.ai to optimize their research protocols, scientists can accelerate the pace of discovery and drive progress in these important areas of study.

More about "Lamin B1"

Lamin B1, also known as LMNB1, is a type of nuclear lamina protein that plays a crucial role in regulating gene expression, chromatin organization, and cellular differentiation.
This essential component of the cell nucleus is responsible for maintaining its structural integrity and is involved in various cellular processes, including DNA repair, transcription, and cell cycle regulation.
Lamin B1 has been extensively studied in the context of several diseases, such as premature aging disorders, neurodegenerative diseases, and cancer.
Researchers can optimize their Lamin B1 studies by utilizing the PubCompare.ai platform, which helps locate relevant protocols from literature, preprints, and patents, and provides AI-driven comparisons to identify the best protocols and products.
This can enhance the reproducibility and accuracy of Lamin B1 research, leading to advancements in our understanding of this important protein and its implications for human health.
PubCompare.ai's powerful tools can elevate your Lamin B1 studies by providing access to a wealth of information, including Ab16048 (a specific Lamin B1 antibody), PVDF membranes (a common substrate for Western blotting), and NE-PER Nuclear and Cytoplasmic Extraction Reagents (a tool for isolating nuclear and cytoplasmic fractions).
By incorporating these resources and leveraging the platform's AI-driven comparisons, researchers can optimize their experimental design, improve data reliability, and ultimately contribute to a deeper understanding of Lamin B1 and its role in human health and disease.
One common method for studying Lamin B1 is Western blotting, which allows for the detection and quantification of Lamin B1 protein levels.
The use of Anti-Lamin B1 antibodies, such as Ab16048, and β-actin as a loading control can provide valuable insights into Lamin B1 expression and its relationship with other cellular processes.
By combining these techniques with the resources and tools offered by PubCompare.ai, researchers can elevate their Lamin B1 studies and drive forward the field of nuclear lamina research.