The reagents used for cell treatment were Lipofectamine 3000 (L3000015, Invitrogen), ascorbic acid (A4544, Sigma-Aldrich), sodium β-glycerophosphate (G9422, Sigma-Aldrich), and dexamethasone (D4902, Sigma-Aldrich). Primary antibodies included PER2 (NBP2-24616, Novus Biologicals), BMAL1 (NB100-2288, Novus Biologicals), PPARγ (A0270, ABclonal Technology, China), AKT1-Ser473 (AP0140, ABclonal Technology, China), β-catenin-Ser552 (AP0579, ABclonal Technology, China), AKT1 (A11016, ABclonal Technology, China), AMELX (ab153915, Abcam), β-catenin (A11343, ABclonal Technology, China), LAMIN B1 (A1910, ABclonal Technology, China), β-tubulin (PMK081M, BioPM, China), GAPDH (PMK042M, BioPM, China), CK14 (MA5-11599, ThermoFisher Scientific), F-actin (ab205, Abcam), and E-cadherin (sc-8426, Santa Cruz).
Lamin b1
Manufactured by ABclonal
Sourced in China
Lamin B1 is a nuclear envelope protein that is a structural component of the nuclear lamina. It helps maintain the structural integrity of the cell nucleus.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Ab153915, by Abcam (1 mentions)
Nbp2 24616, by Novus Biologicals (1 mentions)
β catenin, by ABclonal (1 mentions)
Bmal1, by Novus Biologicals (1 mentions)
Pparγ, by ABclonal (1 mentions)
Ma5 11599, by Thermo Fisher Scientific (1 mentions)
Nb100 2288, by Novus Biologicals (1 mentions)
G9422, by Merck Group (1 mentions)
D4902, by Merck Group (1 mentions)
F actin, by Abcam (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
Anti p p53, by Abcam (1 mentions)
Vdac1, by Abcam (1 mentions)
Anti atpb, by Proteintech (1 mentions)
Enhanced chemiluminescence kit, by Thermo Fisher Scientific (1 mentions)
Anti caspase 9, by Abcam (1 mentions)
Anti sdhb, by Abcam (1 mentions)
Anti bcl xl, by Abcam (1 mentions)
6 protocols using lamin b1
1
Molecular Mechanisms of Cell Differentiation
2
Immunoblotting Assay for Apoptosis Markers
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An immunoblotting assay was performed as previously described [7 (link), 8 (link)]. The primary antibodies used were anti-cytochrome C (1 : 1000, Abcam, UK), anti-Bcl2 (1 : 500, Genspan, USA), anti-Bax (1 : 1000, Abcam, UK), anti-SDHB (1 : 1000, Abcam, UK), anti-parp (1 : 1000, CST, USA), anti-Puma (1 : 500, Santa Cruz, USA), anti-ATPB (1 : 1000, Proteintech, USA), anti-caspase 9 (1 : 1000, Abcam, UK), anti-P53 (1 : 1000, CST, USA), anti-P-p53 (1 : 1000, Abcam, UK), and anti-Bcl-xL (1 : 1000, Abcam, UK). Lamin B1 (1 : 1000, ABclonal, China), VDAC1 (1 : 1000, Abcam, UK), and Gapdh (1 : 2000, Proteintech, USA) were used as the controls. The antigens on the blots were visualized by using the enhanced chemiluminescence kit (Thermo Fisher Scientific, USA).
3
Immunoprecipitation and Western Blot Analysis
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The hnRNPA1 (Cat# A7491), YY1 (Cat# A19569), β-tubulin (Cat# A14991), Lamin B1 (Cat# A16909) antibodies and secondary antibodies were purchased from ABclonal (Wuhan, China) and the FEN1 antibody (Cat# GTX101777) was obtained from GeneTex (Irvine, CA, USA). Phospho-Histone H2A.X (Ser139) antibody (γH2AX) (Cat# 2577) was ordered from Cell Signaling Technology (Danvers, MA, USA). The DNA/RNA oligonucleotides and modified sequences are listed in the Supplementary Materials and were all purchased from Sangon Biotech, (Shanghai, China). Dynabeads M280 streptavidin magnetic beads were obtained from Thermo Fisher Scientific (Waltham, MA, USA).
4
NF-κB Signaling Pathway Analysis
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Tissue and cell extracts were isolated with RIPA buffer (Thermo Fisher, Cat# 89901) supplemented with PhosStop phosphatase inhibitors (Sigma Aldrich, Cat# 4906845001) and cOmpleteTM protease inhibitor mixture (Sigma Aldrich, Cat# 11836153001). Protein extracts were resolved by SDS-PAGE. Blots were incubated overnight at 4°C with antibodies against the following proteins (source, catalog number, and dilution of the antibody are given in parentheses): NF-κB p65 (Cell Signaling Technology, # 6956, 1:500), p-NF-κB p65 (S536) (Cell Signaling Technology, # 3033, 1:500), NLRP3 (Novus, # NBP2-12446, 1:500), Caspase-1 (ABclonal, # A0964, 1:1000), IκBα (Cell Signaling Technology, # 4814, 1:1000), and Lamin B1(ABclonal, # A16909, 1; 500). After washing with TBS-T buffer, blots were incubated with fluorescent (IRDye800 [1:3,000] or IRDye680 [1:3,000], LI-COR Biosciences) conjugated secondary antibodies. The bands were visualized by a Bio-Rad ChemiDoc imaging system (Bio-Rad Laboratories, Inc, USA).
5
Western Blot Analysis of ESCC Proteins
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Total proteins were obtained from ESCC cells using RIPA extraction reagent (Beyotime) supplemented with PMSF protease inhibitor. The concentration was determined by the BCA method. The proteins were separated by SDS-PAGE, and then transferred to PVDF membranes (Millipore). The primary antibodies were incubated with PVDF membrane at room temperature for 2 h or at 4 °C overnight after blocking with skimmed milk. Subsequently, the membranes were exposed to the horseradish peroxidase-labeled secondary antibody for 2 h at room temperature and the chemiluminescent reagent (Millipore) was utilized to detect the protein bands. Primary antibodies against the following proteins were applied: NRSN2 (A14425, ABclonal), Lamin B1 (A11495, ABclonal), GAPDH (#5174, Cell Signaling Technology). Anti-rabbit IgG (#7074, Cell Signaling Technology) was employed as a secondary antibody.
6
Antibody Sources for Western Blot and ChIP
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The sources of antibodies against the following proteins were: β-actin (A1978, 1:10,000 for WB) from Sigma; BMI1 (A0147, 1:1,000 for WB), Lamin B1 (A11495, 1:1000 for WB), MCP-1 (A7277, 1:1,000 for WB), p-p52-s392 (AP1137, 1:1,000 for WB) and p21 (A1483, 1:1,000 for WB) from ABclonal. RNA polymerase II (ab264350, 1:200 for ChIP), H3K4me3 (ab8580, 1:200 for ChIP) and H2AK119Ub (ab195467, 1:200 for ChIP) from abcam.
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