I am running mouse amyloid beta 42 ELISA using the kit from Invitrogen. According the manufacturers guideline, the samples should be diluted so that the concentration of guanidine in final solution is
Yes, some users have reported experiencing standard curve depression when using the Mouse Amyloid Beta 42 ELISA Kit from Invitrogen. This issue has been discussed on various scientific discussion forums and social media platforms.
One possible cause of standard curve depression in this assay is the high concentration of guanidine in the homogenization buffer. As you mentioned, the homogenization buffer provided with the kit contains 5M guanidine, which can interfere with the binding of the antibodies to the amyloid beta peptides and result in reduced signal intensity.
To address this issue, some users have suggested several strategies:
Saif Wahid Thank you very much for your valuable suggestions! I do have some follow up question of your suggestions
Suggestions 1 and 2.. They kit does not come with a homogenization buffer, the used have to prepare the buffer. Did you mean to say homogenize with less concentrated guanidine buffer instead of the suggested 5M concentration? Because after homogenization, samples were diluted to final concentration of 0.08M guanidine in sample
As for suggestion 3, I ran another trial with BSA in the dilution buffer however with no success. I will try another run with BSA in the homogenization buffer. What concentration of BSA should I use? If you could suggest, that would be big help! Thanks!
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