I would like to extract proteins from soybean callus, then I am looking for a protocol to obtain high amounts of protein. I would like to quantify a protein using ELISA methodology after the extraction.
follow this
Article Protein extraction for callus and node cultures of Vanilla p...
You should simply cut and chop soybean callus in to small pieces, homogenize it in 10% TAC, centrifuge it in cold at 10000X G, and resolubilize the pellet and again homogenize and centrifuge it at same speed, please pool both supernatants, and now finally spin both at 25,000 X G for membrane bound proteins, take out supernatant, now use this material for protein estimation with ELISA technique.
To purify protein extracts, use alcohol precipitation method in Wessel, D., Flügge, U.I., 1984. Please find it in this paper in the reference: http://www.ncbi.nlm.nih.gov/pubmed/25578509
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA