Lamin b sc 6216

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Lamin B (sc-6216) is a primary antibody product offered by Santa Cruz Biotechnology. It is used to detect the presence and distribution of Lamin B, a type of lamin protein, in various cell and tissue samples. This product is suitable for applications such as Western blotting, immunohistochemistry, and immunofluorescence.

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Lab products found in correlation

Bay 11 7082 b5556, by Merck Group (1 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Cell culture media, by Thermo Fisher Scientific (1 mentions) Anti flag f3165, by Merck Group (1 mentions) Anti ha hrp, by Roche (1 mentions) Bax 2772, by Cell Signaling Technology (1 mentions) Nf κb p65, by Merck Group (1 mentions) P mek, by Cell Signaling Technology (1 mentions) C caspase 3, by Cell Signaling Technology (1 mentions) P c jun, by Cell Signaling Technology (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) P erk, by Cell Signaling Technology (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Lps l3129, by Merck Group (1 mentions) Anti iba1 antibody 019 19741, by Fujifilm (1 mentions) Immobilon p pvdf membrane, by Merck Group (1 mentions) Anti rabbit antibody, by Cell Signaling Technology (1 mentions)

13 protocols using lamin b sc 6216

Glutaminase Inhibition Mechanism Study

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Phorbol 12-myristate 13-acetate (PMA) (P8139) and BAY 11-7082 (B5556) were bought from Sigma-Aldrich (St. Louis, MO, USA). Bis-2-(5-phenyl acetamido-1,3,4-thiadiazol-2-yl)ethyl sulfide (BPTES) (S7753) was purchased from Selleck Chemicals (Houston, TX, USA).
Mouse anti-GLS1 (ab60709), rabbit anti-GLS1 (ab156876), and rabbit anti-p65 (ab16502) antibodies were bought from Abcam (Cambridge, UK); c-Myc (sc-40), β-actin (sc-130300), and Lamin B (sc-6216) antibodies were purchased from Santa Cruz (Dallas, TX, USA).

Dong M., Miao L., Zhang F., Li S., Han J., Yu R, & Qie S. (2018). Nuclear factor-κB p65 regulates glutaminase 1 expression in human hepatocellular carcinoma. OncoTargets and therapy, 11, 3721-3729.

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Cell Signaling Pathway Analysis

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Cell culture media, gentamicin, penicillin, and L-glutamine were all obtained from Invitrogen (Grand Island, NY). Fetal bovine serum (FBS) was from Gemini Bio-Products (West Sacramento, CA). Tris, NaCl, and SDS for molecular biology and buffer preparation were purchased from Sigma-Aldrich (St. Louis, MO). Antibodies to detect TRAF1 (sc-1830), β-Actin (sc-47778), BRAF (sc-166), GAPDH (sc-25778), NF-κB (p50) (sc-7178), Bcl-2 (sc-7382) and Lamin B (sc-6216) were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Anti-V5 (R96025), V5-HRP (R961-25) and His-HRP (R94125) were from Invitrogen. Anti-HA (901503) was obtained from Covance (Emeryville, CA) and anti-HA-HRP (12013819001) was purchased from Roche (Indianapolis, IN). Anti-Flag (F-3165) was from Millipore. p-BRAF (#2696), NF-κB (p65) (#3034), p-MEK (#9121), MEK (#9122), p-ERK (#9101), ERK (#9102), p-c-Jun (#2361), c-Jun (#9165), p-c-Fos (#5348), c-Fos (#2250), caspase-3 (#9662), c-caspase-3 (#9661), PARP (#9542), c-PARP (#9541) and Bax (#2772) antibodies were purchased from Cell Signaling Technology (Danvers, MA).

Wang Q., Gao G., Zhang T., Yao K., Chen H., Park M.H., Yamamoto H., Wang K., Ma W., Malakhova M., Bode A.M, & Dong Z. (2018). TRAF1 is critical for regulating the BRAF/MEK/ERK pathway in non-small cell lung carcinogenesis. Cancer research, 78(14), 3982-3994.

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Signaling Pathways Analysis Protocol

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Antibodies for phospho-JNK (4668) and phospho-p38 (9215) were purchased from Cell Signaling Technology Inc. The anti-phospho-ERK1/2 (sc-7383), Peli1 (sc-271065), Hsp60 (sc-13115), iNOS (sc-7271) and Lamin B (sc-6216) antibodies were from Santa Cruz. The anti-TH antibody (MAB318) was from Chemicon, anti-Iba1 antibody (019-19741) was from WAKO, and anti-GFAP antibody (G3893) were from Sigma-Aldrich. LPS (L3129) was from Sigma-Aldrich. The cycloheximide (CHX) (HY12320) was from MCE. Murine IL-1β (211-11B) and TNF (315-01 A) were from Peprotech.

Dai D., Yuan J., Wang Y., Xu J., Mao C, & Xiao Y. (2019). Peli1 controls the survival of dopaminergic neurons through modulating microglia-mediated neuroinflammation. Scientific Reports, 9, 8034.

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Western Blot Analysis of Cellular Signaling

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Whole cell lysates and nuclear extracts were loaded on 7% or 12% SDS–polyacrylamide gels, respectively. Electrophoresis was conducted at a constant voltage of 25 V/cm, followed by transfer of the proteins onto an Immobilon-P PVDF membrane (Millipore, Billerica, MA, USA). Membranes were blocked in a solution of 5% phosphoBLOCKER (Cell Biolabs, San Diego, CA, USA) or 5% non-fat milk in PBS-Tween 0.05% for 45 min. Primary antibodies recognizing ERK1/2 (#9107), phospho-ERK1/2 (Thr202/Tyr204, #4377), AKT (#2920), phospho-AKT (Thr308, #4056), phospho-AKT (Ser473, #9271), phospho-H2AX (Ser139, #2577) and PARP (#9542) were purchased from Cell Signaling Technology (Danvers, MA, USA), Lamin B (#sc-6216) and GAPDH (#sc-47724) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Secondary HRP-conjugated anti-mouse (#sc-516102) and anti-goat (#sc-2768) antibodies were from Santa Cruz Biotechnology and anti-rabbit antibodies (#7074) were from Cell Signaling. For protein visualization, membranes were incubated with Pierce^® ECL Western Blotting Substrate (Pierce, Rockford, IL, USA) for 1 min and visualized using the ChemiDoc Imaging System (BioRad, Hercules, CA, USA). ImageJ software was used for densitometric analysis.

Osrodek M., Rozanski M, & Czyz M. (2020). Insulin Reduces the Efficacy of Vemurafenib and Trametinib in Melanoma Cells. Cancer Management and Research, 12, 7231-7250.

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Immunofluorescence Analysis of Lamin-B

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The immunofluorescence analysis was performed according to the procedure previously reported [8 (link)] with brief addition for the inner nuclear membrane Lamin-B protein localization; anti-Lamin B antibody (Lamin B sc-6216 SANTA CRUZ BIOTECHNOLOGY, INC), the secondary antibody red signal-Alexa Fluor 546. Hoechst 33342 (Blue signal after DNA binding). Z-axis series were obtained using a Leica SP8 confocal microscopy.

Salazar C., Yañez O., Elorza A.A., Cortes N., García-Beltrán O., Tiznado W, & Ruiz L.M. (2020). Biosystem Analysis of the Hypoxia Inducible Domain Family Member 2A: Implications in Cancer Biology. Genes, 11(2), 206.

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Antibodies for Nuclear Factor Signaling

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Purified polyclonal antibodies against human Bcl3 (sc-185), Bcl2 (sc-492), NFκB p65 (sc-372X), NFκB p50 (sc-7178X), cRel (sc-71X), RelB (sc-226X), NFκB p52 (sc-848X), and lamin B (sc-6216) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Purified polyclonal antibody against lactate dehydrogenase (LDH; 20-LG22) was from Fitzgerald Industries International (Concord, MA, USA), and actin antibody was from Sigma (St. Louis, MO, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit, anti-mouse and anti-goat secondary antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Bortezomib was purchased from ChemieTek (Indianapolis, IN, USA). Recombinant human IL-8 and IL-17 proteins were from R&D (Minneapolis, MN, USA). All other reagents were molecular biology grade and were from Sigma (St. Louis, MO).

Chang T.P, & Vancurova I. (2014). Bcl3 regulates pro-survival and pro-inflammatory gene expression in cutaneous T-cell lymphoma. Biochimica et biophysica acta, 1843(11), 2620-2630.

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Immortalized Human and Rat HSC Cell Lines

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The immortalized human HSC line LX-2 and rat HSC line HSC-T6 were used (American Type Cell Culture, Manassas, VA). Both cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies Corporation, Grand Island, NY) with a high glucose concentration (4.5 g/L) supplemented with 5% fetal bovine serum (FBS), 100U/ml penicillin G and 100 μg/ml streptomycin (Gibco BRL, Gaithersburg, MD). Human hepatic hepatocytes C3A were also used and cultured in Eagle's Minimum Essential Medium (American Type Cell Culture, Manassas, VA) supplemented with 10% FBS. All cells were incubated at 37°C in a humidified atmosphere containing 5% CO₂. The following antibodies were used: Alpha-smooth muscle actin (α-SMA) (A5228) and SAA2 (SAB1401350) (Sigma-Aldrich, St. Louis, MO); Fibronectin (sc-6952), STAT3 (sc-482), SOCS3 (sc-9023), Lamin-B (sc-6216), and alpha-Tubulin (sc-8035)(Santa Cruz Biotechnology, Santa Cruz, CA); phospho-STAT3 (Y705) (ab34716)(Abcam, Cambridge, MA); Collagen Type I (600-401-103) (Rockland Immunochemicals, Gilbertsville, PA); GAPDH (5174), Smad2/3 (8685) and phospho-Smad2/3 (8828) (Cell Signaling Technology, Danvers, MA; 5174); and Topoisomerase Iiβ (611492) (BD Biosciences, Franklin Lakes, NJ). The STAT3 inhibitors S31-201 (CAS501919-59-1) (Santa Cruz Biotechnology, Santa Cruz, CA) and FLLL32 (530153) (Millipore, Darmstadt, Germany) were used.

Lopez O.N., Bohanon F.J., Wang X., Ye N., Corsello T., Rojas-Khalil Y., Chen H., Chen H., Zhou J, & Radhakrishnan R.S. (2016). STAT3 Inhibition Suppresses Hepatic Stellate Cell Fibrogenesis: HJC0123, a Potential Therapeutic Agent for Liver Fibrosis. RSC advances, 6(102), 100652-100663.

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TGFβ3 Signaling Pathway Regulation

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Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA). PD98059, SB202190, SP600125, LY294002, SIS3, and actinomycin D were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). H-89 and GF109203X were obtained from Biomol (Farmingdale, NY, USA). The antibody (Ab) generated against phospho-Smad2/3 (#33102) was obtained from Cell Signaling Technology (Danvers, MA, USA). The Abs raised against total Smad2/3 (sc-11769) and lamin B (sc-6216) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). The Abs raised against total Smad2/3 (sc-11769) and lamin B (sc-6216) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The Ab raised against α-tubulin (GTX628802) was purchased from GeneTex (Irvine, CA, USA). TGFβ3 was dissolved in 4mM HCl-0.1% bovine serum albumin, which was used as a vehicle in this study.

Lai T.H., Chen H.T., Wu P.H, & Wu W.B. (2024). The Presence of TGFβ3 in Human Ovarian Intrafollicular Fluid and Its Involvement in Thromboxane Generation in Follicular Granulosa Cells through a Canonical TGFβRI, Smad2/3 Signaling Pathway and COX-2 Induction. International Journal of Molecular Sciences, 25(10), 5558.

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Comprehensive Protein Profiling of Cell Signaling

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Whole cell lysates were prepared using RIPA buffer containing phosphatase inhibitor and protease inhibitor cocktail (Roche, Basel, Switzerland). Following standard western blotting procedure, proteins were blotted onto PVDF membrane (Millipore, Bilerica, MA, USA) and recognized by the following antibodies: SRSF3 (sc-13510), α-tubulin (sc-69969), and lamin B (sc-6216) were from Santa Cruz Biotechnology; Sp1 (#5931), IKKα (#2682), IKKβ (#2370); p-IKKα/β (#2697), NF-κB p65 (#8242), p-NF-κB p65 (#3033), Cyclin D1 (#2926), and Slug (#9585) were from Cell Signaling Technology (Danvers, MA, USA); IκB-β (A301-828A) was from Bethyl Laboratories (Montgomery, TX, USA); HIF1-α (610958) was from BD Bioscience; NKIRAS2 (ab57303), GAPDH (ab8245), and β-actin (ab8226) were from Abcam (Cambridge, UK).

Kim H.R., Shin C.H., Lee H., Choi K.H., Nam D.H., Ohn T, & Kim H.H. (2016). MicroRNA-1908-5p contributes to the oncogenic function of the splicing factor SRSF3. Oncotarget, 8(5), 8342-8355.

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PBrP Modulation of TGF-β Signaling

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To examine the effect of PBrP on TGF-β-induced Smad2/3 phosphorylation, Mv1Lu and A549 cells were incubated with PBrP at various concentrations or for various periods and then stimulated with 100 pM of TGF-β for 30 min. To monitor EMT protein expression, the cells were pretreated with different PBrP concentrations for 2 h and then stimulated with TGF-β for 48 h. The cells were lysed with a lysis buffer and standardised using a bicinchoninic acid (BCA) protein assay (Pierce™ BCA Protein Assay Kit 23225). Fifty microgram of this protein was applied to SDS-PAGE and electrotransferred to the polyvinylidene difluoride (PVDF) membrane. Polyclonal antibodies against TβRI (sc-398), TβRII (sc-400), EGFR (sc-373746), β-actin (sc-47778), flotilin-1 (sc-25506), fibronectin (sc-9068), N-cadherin (sc-7939), Lamin B (sc-6216) and vimentin (sc-7557) were obtained from Santa Cruz (Dallas, TX). Rabbit Polyclonal antibodies against pSmad2/3 (#8828), Smad2/3 (#8685), PAI-1 (#11907), Flag tag (#8146), Lamp-1 (sc-9091) and MyoVa (#3402) were purchased from Cell Signalling (Boston, MA). Secondary antibodies conjugated with horseradish peroxidase (Millipore, Darmstadt, Germany) and an enhanced chemiluminescence (ECL) kit (Perkin-Elmer Life Sciences, Waltham, MA, USA) were used to develop immunoblots.

Shih-Wei W., Chih-Ling C., Kao Y.C., Martin R., Knölker H.J., Shiao M.S, & Chen C.L. (2018). Pentabromopseudilin: a myosin V inhibitor suppresses TGF-β activity by recruiting the type II TGF-β receptor to lysosomal degradation. Journal of Enzyme Inhibition and Medicinal Chemistry, 33(1), 920-935.

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