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Dear Sir/Madam 1) We are doing western blot experiments for quantification. I read in the antibody guides that it would not be an advantage using polyclonals in quantification experiments. Is it absolutely not recommended using polyclonals when we are quantifying? 2) Can you please recommend a positive control and a blocking peptide for IL7Ralpha (ab95023) 3) Can you please recommend a blocking peptide for CCR7 (ab103404) 4) What antibody for TLR-4 and CD45 do you recommend? Experiment: Western blot on human blood samples (mostly white blood cells (but contaminated with plasma and red blood cells)). We are going to quantify the bands. We would also appreciate recommendations for positive controls and for blocking peptides fore these antibodies. Kind regards
Asked on Sep 08 2011
Thank you for contacting us. The positive controls for ab95023 are cell lysates of A-431, U2OS, U251 MG, Hela cell etc. Please click the following to see the positive control tissue lysates; http://www.proteinatlas.org/ENSG00000168685 We unfortunately cannot offer blocking peptide for this antibody; the peptide is not commercially available. We can certainly offer a blocking peptide for ab103404; let me know if you are interested in buying this we will then publish this product as special request. TLR4 antibody:- We can recommend ab83444; https://www.abcam.com/tlr4-antibody-ab83444.html The blocking peptide for this is ab93304; https://www.abcam.com/TLR4-peptide-ab93304.html The positive control lysates would be Human, Mouse and Rat small intestine; NIH3T3; EL4 etc. CD45 antibody and peptide would be:- ab10559; https://www.abcam.com/CD45-antibody-ab10559.html and ab17550; https://www.abcam.com/CD45-peptide-ab17550.html The positive control will be Jurkat, KM-H2, or human tonsil tissue lysates. Please also check human tissue Atlas for more information about positive control. Polyclonal antibodies can be used for protein quantifications also however because monoclonal antibodies binds more specifically to particular proteins so monoclonal antibodies are recommended more than polyclonal antibodies. The choice antibodies are depends on the type of application used e.g. in sandwich ELISA monoclonal is best. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Sep 08 2011