I am using an fibrinogen antibody to detect Fibrin degradation products in the plasma clot supernatent. This antibody detects all the fragments of fibrinogen.
I am using indirect ELISA for this, I coat 96 well plate with supernatent in the coating buffer(Carbonate) for overnight and next day wash the wells by adding TBSt and inverting the plate 3 times, blocking for 1 hr with 5 % BSA, then incubation with primary antibody for 4 hours and same washing 3 times, followed by secondary HRP conjugated antibody incubation for 2 hrs followed by washing and then adding TMB substrate , incubate for 15 min and adding 1N H2SO4 as stopping solution.
I am using Fibrinogen as my standard. My problem is I have same color in all the wells, I am gussing its the problem of washing so i tried washing it with pipetting out the solution. Butit didnt work, Is there somebody who can suggest me a better way???
Thank you very much... Seeking for help..
Dear Manovriti,
consult the authors to these papers here on RG:
Article The clinical significance of thrombin-antithrombin III compl...
Article New method for determining thrombin-clottable fibrinogen
Article Fibrin degradation during sonothrombolysis – Effect of ultra...
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