β-2-Microglobulin Antibody (BBM.1): sc-13565

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:100) -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 (1:100) -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

Tissue for western blotting was obtained from the New York Brain Bank frozen, and then rapidly homogenated at 4 °C in lysis buffer containing 50 mM of Tris-HCl, 150 mM of NaCl, 5 mM of EDTA and 1% of Triton-Tx. Protein concentration of the total homogenate was measured using the BCA assay (Thermo Scientific). 60 μg of protein per lane were run on a 10% polyacrylamide gel for HLA antibody and on a 15% polyacrylamide gel for β2m antibody. Proteins were transferred to PVDF membranes (Immobilon, Millipore Corporation, Bedford, MA, USA). After blocking in 5 % dry milk, primary antibodies were applied overnight at 4 °C, then rinsed 3 times in Tris-buffered saline containing 0.1% Tween-20 (Fisher Scientific). Peroxidase-labeled secondary antibodies were applied for 1 h at RT, blots rinsed in Tris-buffered saline-Tween as before, developed with enhanced chemiluminescence (Immobilon HRP substrate reagents, Millipore), and then exposed to film (Crystalgen). HLA (1:100, from Santa Cruz Biotechnologies) and β2m (1:100, from Abnova) primary antibodies were used. Actin (1:500,000, from Sigma Aldrich) primary antibody was used as a loading control.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for β-2-Microglobulin Antibody (BBM.1): sc-13565 below.

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