Recombinant Anti-IKK gamma/NEMO antibody [EPR16629] (ab178872)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Rabbit (1:5000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Rabbit (1:5000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

For Western blotting, samples were scraped from 6-well plates in 50 uL RIPA buffer ((0.15 M NaCl, 50 mM Tris pH 7.4, 1% deoxycholic acid, 1% Triton-X100, 0.1% SDS, 1mM EDTA, supplemented with 1× Proteoblock protease inhibitor cocktail). Samples (50ug protein) were run on standard denaturing SDS-PAGE, and proteins were transferred to nitrocellulose (NC) at 400 mA for 80 minutes at room temperature or 12 V overnight at 4 °C. NC membranes were blocked in Odyssey Blocking Buffer (Licor, catalog # 927-40100) and then the primary antibody, rabbit anti-G6PD (Bethyl catalog # A300-404A), was used at 1:1000. Goat anti-actin (Santa Cruz sc-1616) was used as a loading control at 1:100. Secondary infrared-dye conjugated antibodies were used at 1:10,000–20,000 dilutions. NC membranes were scanned on an Odyssey infrared scanner. For NEMO (IKKγ) Western blotting, we used Abcam rabbit monoclonal antibody # ab178872 at a 1:5000 dilution.

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Manufacturer protocol

Download the product protocol from Abcam for Recombinant Anti-IKK gamma/NEMO antibody [EPR16629] (ab178872) below.

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