Protocol Online logo
Top : New Forum Archives (2009-): : ELISA and Immunoassay

ELISA Standard curves and sample concentration - (Aug/30/2009 )

Hi all, I using ELISA commercial kit. At first, i ran my first ELISA run using duplicate samples, but later on i found some big standard variation so thought might run another plate using same sample (i thought i can use my second plate result to replace one of the first plate's results). However, because the kit required us to run Standard everytime we run the assay, and ended up my second plate result of same sample had different concentration. The absorbance value for the sample in Run 1 and Run 2 are almost the same, but the standard curve is different. Do you think i allow to check my Run 2 sample concentration using the Run 1 standard curve? Or what would you advice? Thanks

-wntiong-

wntiong on Aug 31 2009, 03:14 AM said:

Do you think i allow to check my Run 2 sample concentration using the Run 1 standard curve?


Itís tempting isnít it :( but afraid not, unless you have a good reason like the pipette tips fell off (Or possibly if you had run several plates and only one was out).
If you are mainly interested in comparing between samples rather than accurate quantitation the standard curve is not going to matter so much as long as the trends are the same for both plates.

-DRT-