Hi all.
I am working on animal colitis model (rats) experiments and trying to find TNF-alpha and IL-6 amounts in colon tissue samples. After homogenization and centrifugation, I used the Elisa Kit method and used 100 ul samples. Taking 100 ul supernatant from samples is the correct way or we need first to find total protein and then do analysis. I am wondering then how to make the final calculations, means take fix amount of protein for all samples or volume ...Please comments
Hi, you should do a simple bradford protein assay simultaneous to your ELISA to determine protein concentration (mg/ml). Then when done with assay, divide the cytokine concentration (ex. pg/ml) per mg of protein per tissue sample. This will standardize it to ex. pg cytokine per mg protein
Hi, you should do a simple bradford protein assay simultaneous to your ELISA to determine protein concentration (mg/ml). Then when done with assay, divide the cytokine concentration (ex. pg/ml) per mg of protein per tissue sample. This will standardize it to ex. pg cytokine per mg protein
Another option is to weigh the tissue before homogenization and then express it per mg of tissue.
I am totally agree with Gerard Moloney, you should weigh your tissue before make any homogenization of tissue. aslo check which type of buffer or solution your kit require for homogenization of tissue,
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