Hi,
I have been doing HBD-3 Elisa with highest sample concentration 50-50 dilution. I heard that 50-50 is better than adding 100uL of your sample, because you take into account the diluent as well and the blank is not subtracted from your sample. I was told that if a sample is 0 in 50-50 dilution it would be 0 in 100uL as well so there is no point.
Now I want to do IL-8 Elisa and I am looking for not 0 values. Shall I do 50-50 dilution, would that be enough, or would 100uL have more chances of giving me positive values?
(Sensitivity 8 pg/mL Standard Range 15.6-1,000 pg/mL)
Thank you in advance!
Best wishes,
Virginia
It depends on the purity of sample used. I think wide range of standarad will make a problem. firstly try to make check board titeration to get the best dilution.
We routinely do undiluted samples (neat, no 50:50 dilution), esp. if we expect the signal to be super low. The only time we do 50:50 when looking at low signals is if we have very little sample left.
You are right in that if the signal is below LOQ in 50:50 dilution, chances are it will be below LOQ in the neat sample. However, if it is 8 pg/mL in 50:50 dilution, you should have quantifiable amount in your neat (around 16 pg/mL). So it does help. There is absolutely no reason why you can't use neat samples.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA