Immunoprecipitation abreview for Anti-TRPM8 antibody

Immuno-precipitation step
Protein A
Human Cell lysate - whole cell (Hek cells transfected with human TRPM8)
Hek cells transfected with human TRPM8
Total protein in input
800 µg

Other product details

6 µg/mg lysate
Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C

Western Blot antibody

Abcam antibody:Anti-TRPM8 antibody

Additional data

Additional Notes
Cells were washed in ice-cold PBS solution and lysed in HEPES-CHAPS buffer (2% CHAPS in 50 mM Hepes, 200 mM NaCl pH 7.5), and a mixture of protease inhibitors (Roche) for 30 min on ice. Immunoprecipitation was performed by incubation with anti-TRPM8 antibody ab3243 (6 ´g/mg lysate) at 4¯C, overnight. Protein A-Sepharose beads (Sigma) were washed and equilibrated in 50 mM Hepes, 200 mM NaCl pH 7.5 buffer. Western Blot Assay: Immunoprecipitates and whole cell lysates were separated on 8% polyacrylamide gels, followed by transfer onto PVDF membranes. The membrane was processed for chemiluminescence detection using Western Blotting Luminol Reagent (Santa Cruz Biotechnology, Inc.). Image: First lane: IP, Second lane: whole cell lysate.

Miss. Larisa Maier

Verified customer

Submitted Sep 17 2013

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