Formulations of Commonly Used Buffers and Media:
These are the formulations of buffers we have successfully used over the years. If they are not the same as other formulations, the differences are probably not significant.
Dilute to 1X with sterile distilled water and use to remove adherent cells from tissue culture dishes. To do this, aspirate off culture media and replace with 1X solution. Let cells sit in the incubator at 37°C, and then monitor cells with microscope: generally they are no longer adherent after as little as 5 minutes. Collect cells by centrifugation, replace citrate saline with regular culture media, and replate cells. Cheap and Efficient!
Mounting Media for Immunofluorescence Microscopy:
Phosphate Buffered Saline:
1X PBS is 1.47mM KH2PO4, 10mM Na2HPO4, 2.7mM KCl, 137mM NaCl pH=7.4. It is a more or less physiological buffer which living mammalian cells can tolerate at least for a short time. It is frequently used to wash cells prior to protein extraction or immunostaining. It is also useful for antibody incubations in immunocytochemical staining of cells in tissue culture and sections. One caveat is that phosphatase based detection systems are inhibited by high concentrations of phosphate, the phosphatase reaction product, so you need to do a wash in some sort of phosphate free buffer before you use these. For antibody staining and washing you can also add 0.1% Tween 20 (Polyoxyethylene 20-sorbitol monolaurate) or 0.1% Triton X-100 non-ionic detergents to reduce background staining.
Tris Buffered EDTA (TBE):
Tris Buffered Saline:
1X TBS is 10 mM Tris/HCl, 150 mM NaCl, pH=7.5. We use TBS routinely for ELISA, immunoblots etc. Can also add 0.1% Tween 20 or 0.1% Triton X-100 non-ionic detergents to final concentration of 0.1% reduce background.
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