Question (30079) | Human Histone H3 (phospho S10) peptide (ab11477)

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Question

  I need your help, in my blocking peptide experiment. Would you please answer to my questions:   1-      How much antibody “ab5176” should I add to peptide “ab11477”? 2-      Should I first aliquot  both peptide and antibody? In what kind of buffer? 3-      What kind of secondary antibody is suitable?    

Answer

We generally suggest a 200fold molar excess of the peptide. This usually means ca 2ug peptide per 1 ug antibody. This can be increased to up to 500fold molar excess of the peptide if necessary. I suggest to use the same buffer that is used for the incubation steps with the antibody or just PBS Tween (0.1%) We also generally suggest to aliquot all antibodies and peptides to avoid contamination and to store them according to the instructions on the datasheet. A suitable secondary antibody for ab11477 is an anti rabbit antibody that is conjugated to HRP: Click here (or use the following: https://www.abcam.com/index.html?datasheet=6721). Click here (or use the following: https://www.abcam.com/index.html?datasheet=6802). Click here (or use the following: https://www.abcam.com/index.html?datasheet=6795).

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