Hi everyone,
I am interested to know if anyone has had any experience working with HUVEC cell lysates and running them on ELISA's? We seed our HUVEC on a gelatin matrix- could this impact our ELISA results? If anyone has any advice on using cell lysates in general as ELISA samples (or more specifically HUVEC!), this is greatly appreciated!
Many thanks! :)
The gelatin matrix can be used as a blank with the appropriate buffer. this can be compared with the black of HUVEC cell lysates...
These two blanks can be spectrophotometrically also compared over various wavelengths. As a exclusive testing, so see if the peaks are changing anywhere.
ELISA has issues as exceptions...
i think the face lotion being applied by a technician was interferring in T3/T4 assay. the organic solvent of face solotion, was in the hand as the individual using the pipetted.
As such no experience of HUVEC cell lysates.
Daisy Jones . Are you coating HUVECS on plates coated with gelatin or onto plates that have solid gelatin gel?
I have experience in the former and there should be no problem.
Steingrimur Stefansson Hi,
what is the difference between th coating and the solid gelatin gel please, and how do you procede for the caoting?
Thank you
Hi Nait Eldjoudi Amina . For gelatin coating you use 5-10 ug/ml gelatin to cover the TC plates with the protein for ~1 hour at 37oC and then wash the excess gelatin off.
For solid gelatin gels, you use at least 3% warm gelatin and let it congeal as a film on the TC plates.
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