Hello,
I've obtained a protein pellet from my bovine mammary gland tissue samples and would like to use it for ELISA tests. I'm not sure what buffer/water/PBS or other solution should I use to resuspend the pellet. I received it from microRNA extraction (NucleoSpin miRNA kit, Macherey-Nagel) and they recommend to use SDS-containing solution, as Laemmli buffer and incubate it in 90oC but will it be ok for ELISA and not for SDS-PAGE? If not what are your recommendations for resuspending the protein pellet derived from tissue like mine? Also, should I wash the pellet first with 50% ethanol, before redissovling?
Thank you very much in advance!
Are you going to use a sandwich ELISA or bind the protein extract directly to the plate? If the latter, you may be able to dissolve the pellet directly into the binding buffer, which is a mildly alkaline buffer consisting of sodium carbonate and sodium bicarbonate. I wouldn't use this for a sandwich ELISA, however, for fear of damaging the capture antibody. For that, a nonionic detergent (e.g. 1% Tween-20)-containing neutral buffer (e.g. PBS) may be the way to go.
SDS will destroy the protein and will prevent the adsorption of the protein (to the plate or to another protein). As mentioned, try dissolving it first in the carbonate buffer. 50% to wash the pellet from what?
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Inorganic Materials
- Ceramics
- Glass