Has anyone ever extracted protein from tongue tissue to perform ELISA? What kind of lysis buffer did you use?
Hello
I think RIPA buffer is not bad lysis buffer for ELISA, we are using it for lysis of pancreatic tissue, so, just freeze tongue tissue in nitrogen and then make a powder, after that you could easily lyse it in RIPA buffer.
Wish you best in your experiments!!!
Dear Patrick in my opinion choose of an ideal buffer will depend on the localization, in the cell, of antigen you are looking for. If you are studying a membrane antigen then a buffer containing a detergent and preferentially non-ionic as “glycopynaroside” (because excess can be removed by dialysis from solution and it is a soft detergent) can be used. If you are studying a cytosolic molecule then a lysis buffer is indicated, but if the antigen is located on extracellular matrix then you will have to use a buffer containing chaotropic molecules as urea or guanidine, in order to extract ECM constituents. You will find descriptions of detailed buffers in literature. Good look in your experiments. Silvio.
Human Whole Tongue Tissue Lysate
Product Description
Human Whole Tongue Tissue Lysate. (prenatal derived)
100 ug/vial
DV Biologics cell and tissue lysates are prepared from liquid nitrogen fresh frozen samples. Cell and tissue specimens are homogenized in ice-cold modified RIPA buffer in the presence of protease inhibitors (without EDTA). After lysis, the samples are centrifuged to remove tissue and cell debris. The protein concentration is determined using bicinchoninic acid (BCA) assay and BSA as a standard. DV Biologics cell and tissues lysates can be used in SDS-PAGE, Western blotting, immunoprecipitation, ELISA, enzymatic activity analysis, protein-protein interaction and other studies encompassing functional genomics, and comparative analysis of proteins in human models.
I´m agree with Silvio, consider that the buffer is mainly there to solubilize the protein of interest and to keep it stable. I do not think that tongue tissue have to be handle in a different way that other soft tissues.
Best is to hit on pubmed the name of the protein of interest and check on literature how people perform for example protein extraction for western blot.
Best
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