1 reply to this topic

[hhyy]

Hi,

I use the human serum to do the ELISA, but it seems in the coated and non coated wells, i got almost same result, around 1.7; for the another uncoated well, i did not add the serum, the reader is around 0.1; for the other coated wells, i got the reader from 1.3 to 0.5. Any one can help me explain why for the uncoated well also got the high read? and any good suggestions? Thanks.

[protolder]

hhyy, on 25 May 2011 - 11:04 AM, said:

Hi,

I use the human serum to do the ELISA, but it seems in the coated and non coated wells, i got almost same result, around 1.7; for the another uncoated well, i did not add the serum, the reader is around 0.1; for the other coated wells, i got the reader from 1.3 to 0.5. Any one can help me explain why for the uncoated well also got the high read? and any good suggestions? Thanks.

Hola, if you have a high signal is because the first antibody bounds to the surface, so your blocking stage isn´t run well.In uncoated wells,  0.1 could be a background of experiment if the developing time is quite long, but 1.5 shows that the well is bad blocked. So or have you forgotten add blocking agent or it´s isn homogeneous. Remember that the volume of blocking solution has to be greater than coating solution volume to be sure that all the surface and wall of the well is blocked. . Buena suerte