Hello all,
I have IFN-alpha kit, which is having IFN-ALPHA protein as its standard. Can this protein be treated as reagent and used as inhibitory or stimulatory agent in other assays like apoptosis assay?
I have no idea which company kit you are using, but I have found a particular suggestions from several companies regarding to the use of standards. For example eBioscience or Raybiotech the standard are not very stable and they can be used only once if you dissolved them. My suggestion is not to use for other experiments until there is assurance of there stability and other use.
Hi Ashwini,
I would also strongly advise against using standards from ELISA kits in any functional applications. Protein used as standard is not necessarily the full-length active protein but can be a partial sequence, possibly carrying a tag (e.g. His-tag) that was used for purification. So unless the manual specifically states that the standard is biologically active I would refrain from using it in a functional assay.
Best,
Christian
Thank you so much to both of you..I will surely consider your suggestions..
In our lab, we have used the standards from ELSIA kits as stimulants in other assays.
We purchase both ELISA kits and a variety of stimulating agents from the same company (Peprotech). We have found that using proteins purchased as stimulants or borrowing ELISA standards to be used as stimulants, produced similar (expected) cellular responses.
Since the standards for the ELISA kits are always in excess compared to the antibodies, using up the standards as stimulating agents in other assays saves reagents and laboratory costs.
In addition to the chemical structure of IFN-ALPHA protein, you should take into consideration the buffers, pH of the solutions, concentrations, quantity of salts to be used in other kits. Do your pre-experiments and make your decision.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA