Question (32825) | Streptavidin Conjugation Kit (ab102921)

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Question

Hello I bought from you Streptavidin Conjugation kit – ab102921 – and I used 200ul of Ab (conc 500ug/ml) – so I had 100ug in 200ul and I realized that conjugation efficiency was not good (a lot of Ab`s were uncojugated). In recommended amount and volume of Ab for optimal results you suggest to have 100-200ug of Ab in max 100ul. So my question is – what should I do to obtain good efficiency with 200ul of Ab in concentration 500ug/ml Best wishes

Answer

Thank you for your inquiry. I am sorry to hear that ab102921 did not deliver optimal results for you. I recommend to use the kit as instructed with a maximum of 100ul and 100ug of antibody. These kits are optimized for this specific protocol. There is the option of concentrating the antibody in order to be able to use 100ug for the reaction while not exceeding the maximum volume. I can recommend ab102778 for this. Click here (or use the following: https://www.abcam.com/index.html?datasheet=102778). The Easy Link Kits work by targeting 'available' amines on the antibody. They will also label proteins etc that have amines. This is the reason not to use more volume as stated and to use purified antibody. Otherwise the streptavidin will label the proteins in the buffer instead of the antibodies. Many antibodies come in buffers that contain additional protein to keep the antibody stable. I suggest to purify the antibody in this case. I can recommend ab102784 for purification. Click here (or use the following: https://www.abcam.com/index.html?datasheet=102784). I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

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