Question (10357) | Anti-Antithrombin III/ATIII antibody (ab8776)

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Question

Thank you for your help last time. Now I am still looking for two antibodies of your own---ab8776, ab13335, and I have some questions about their applications. I would like to use them in sandwich ELISA experiment to quantify antithrombin in human plasma. But, within these two antibodies, only ab13335 has been tested for ELISA. ab8776 has just been tested for IEP, RID.Therefore I would like to know if I can combine these two antibodies in ELISA and which one I should coat on the microplate. And please send me your protocols and tell me the dilution rate you've used. Sorry for asking so many questions. Your answers will be a great help to me. Please answer me as soon as possible.

Answer

I have received the following protocol for ab13335 in direct ELISA (not sandwich as sandwich ELISA has not been tested with ab13335) which I hope will help you. 1. Coating 1-5 ug/ml per well of Anti-thrombin III on High binding plates. Incubation for overnight at 4C. 2. Blocking with 1% BSA in PBS for 30 minutes. 3. Wash buffer: PBS Tween 0.05%. 4. Primary antibody: this product at 1:1,000-1:2,000 dilutions. Incubation for 120 minutes at room temperature. 5. Secondary antibody: HRP APA mouse IgG at 1:500-1:1,000 dilutions. Incubation for 120 minutes at room temperature. 6. Substrate: OPD Please let me know if this helps. Best of luck with your research

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