Lamin B1 Antibody (C-5): sc-365962

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:500) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:500) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Cells were harvested using RIPA (radio immunoprecipitation assay) buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% Nonidet-P40, 0.5% sodium deoxycholate and 0.5% SDS) supplemented with phosphatase (Na3VO4, 1:100, NaF, 1:200 or phosphatase inhibitor cocktail 2, Sigma, 10 μl ml−1) and protease inhibitors (phenylmethylsulphonyl fluoride in ethanol or protease inhibitor cocktail, Sigma). Proteins were purified by centrifugation at 15,000g at 4 °C for 20 min and then quantified using bicinchoninic acid assay kit (Thermo Fisher Scientific) and resolved on 7.5% Bis-Tris polyacrylamide gels, before being transferred onto polyvinylidene fluoride membranes via electroblotting. Blots were probed with primary antibody overnight followed by horseradish peroxidise (HRP)-conjugated secondary antibodies before being developed via enhanced chemiluminescent substrate and exposure onto CL-XPosure film (Thermo Fisher Scientific). Molecular weight markers (Thermo Fisher Scientific 11832124 or New England BioLab #7712) were run alongside all samples and values are as indicated in both the main and uncropped scans

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Manufacturer protocol

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