Cells were lysed with buffer A (25 mM Hepes, 150 mM NaCl, 1 mM EGTA, 10 mM NaF, 10 mM Napyrophosphate, 5 mM sodium orthovanadate, protease inhibitors cocktail) containing 1 % NP-40 and 10 % glycerol (lysis buffer) at 4 °C. Lysates were incubated with the anti-Fas antibody Apo1.3 (Enzo Life sciences, Farmingdale, New York) or the anti-HA antibody (clone 16B12, Covance, Berkeley, USA) coupled to protein G-sepharose beads (Zymed, San Francisco, USA) for 4 hours. After 4 washes with lysis buffer, beads were eluted with Laemmli buffer at 95°C for 5 min and subjected to immunoblotting analysis.
Anti ha antibody clone 16b12
Manufactured by Fortrea
Sourced in United States
Anti-HA antibody (clone 16B12) is a laboratory reagent used for the detection and purification of proteins tagged with the hemagglutinin (HA) epitope. It is a mouse monoclonal antibody that specifically recognizes the HA tag sequence.
Automatically generated - may contain errors
2 protocols using anti ha antibody clone 16b12
1
Immunoprecipitation and Immunoblotting Analysis
2
Western Blot Analysis of Protein Markers
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Western blot analysis was carried out using the Odyssey Infrared System (LI-COR Biosciences, Lincoln, NE) and anti-β-catenin antibody #9582 (Cell Signaling, Danvers, MA), anti-PIWIL1 antibody ab12337 (Abcam, Cambridge, MA), anti-HA antibody clone 16B12 (Covance, Berkeley, CA), monoclonal beta actin antibody AC-74 (Sigma, St. Louis, MO), and fluorescently-labeled secondary antibodies IRDye 680RD Infrared Dye (LI-COR Biosciences, Lincoln, NE). The experiments were repeated at least 3 times. Protein levels were normalized to actin levels (the loading control) with standard deviations.
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