Blocking Buffer (2X) for Fluorescent Western Blotting

This blocking buffer is specifically designed for western blotting using fluorochrome conjugated antibodies and can be used for membrane blocking and to dilute both primary and secondary antibodies. See www.rockland inc.com for specific protocols. This buffer was prepared using ultra pure reagents dissolved in pharmaceutical grade water (WFI) and consists of a proprietary protein formulation in TRIS buffered saline at pH 7.6 with thimerosal added as an antimicrobial agent.

Western Blot Comparison of 1X and 2X MB-070 Buffer. Lane 1: reduced human NAG1 protein 0.1ug. Lane 2: Prestained Molecular Weight Marker 5uL (p/n MB-210-0500). Lane 3: non-reduced human NAG1 protein 0.1ug. Blocking Buffer: Left blot 1X, Right blot 2X of (p/n MB-070-050) for 30 min at RT. Primary Antibody: rabbit anti-NAG1 biotin conjugated antibody (p/n 209-406-B88) at 1ug/mL overnight at 2-8C. Secondary Antibody: goat anti-rabbit antibody HRP conjugated (p/n 611-103-122) at 1:70,000 for 30 min at RT. Expect: ~14kDa for NAG1.