5 replies to this topic

[miss-pcr]

Hi..
please tell me why my ELISA plate directly gives yellow color after adding the substrate

[pesji]

Which substrate and which secondary antibody do yu use ? Peroxidase labeled or Phosphatase ?

[swanny]

You may not have washed the enzyme off the plate.

[tonix37]

Could be a problem of substract.
When you prepare substracte, can you see background on substracte solution before add it in the wells??

pesji, on Nov 2 2009, 12:56 PM, said:

Which substrate and which secondary antibody do yu use ? Peroxidase labeled or Phosphatase ?

[almost a doctor]

how are you washing your plates before adding the substrate?  most detergents interfere with chromogenic substrates so you need to wash your plates with dH2O a couple of times at the end before developing your plates

[miss-pcr]

Thanks for your all replys,yes my problem is in my substrate and washing procedure