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Dear Sir/Madam, I have a very particular question about Foxp2 antibody ab16046: In the guideline it is said that the antibody gave positive result against HEK293 Whole Cell Lysate. Was the Lysate prepared according to AbCAM protocol "Preparation of lysate from cell culture" (https://www.abcam.com/index.html?pageconfig=resource&rid=11379#A1), using AbCAM RIPA buffer recipe? And that were wild type non transfected cells HEK293, right? That would be very helpful if you could confirm this information for me. Thank you very much in advance, With best regards,
Asked on Nov 21 2011
Thank you for contacting us. In answer to your enquiry, the lysis buffer used to prepare the sample of HEK293 cells was not exactly as from the protocol you have found on our website but was purchased from one of the larger reagent suppliers. The components however are much the same: 150 mM NaCl, 1.0% detergent, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, pH 8.0. The protocol used was also very similar, apart from that incubation with lysis buffer which was performed for 10 minutes on ice followed by sonication. Both a protease inhibitor cocktail and sodium orthavanadate was used in the lysis buffer. 10 µg of whole cell lysate was loaded per well and the primary antibody was used in a 1µg/ml dilution. The FOXP2 protein detected was endogenously expressed, no transfection was preformed. I hope this information has been of help. If you require any further information please do not hesitate to contact us again.
Answered on Nov 21 2011