Anyone can share the protocol to measure mice tissue (spleen) protein concentration for western blot by using Coomassie brilliant blue G 250 and how to calculate from results the amount of sample to load on gel? Thank you.
Hi!
Try the this product and protocol: Bio-Rad Protein Assay Dye Reagent Concentrate.
Protocol ( https://www.bio-rad.com/webroot/web/pdf/lsr/literature/LIT33.pdf ).
The calculation is done based on linear regression equation obtained from a standard curve made with BSA (usually). Interpolate your unknow samples in the curve (line); your sample absorbance must be inside the standard curve range. Dilute or aplly more sample to achieve this.
Best,
Jacó
Hi,
Another option:
Article Fast and Sensitive Colloidal Coomassie G-250 Staining for Pr...
https://www.nacalai.co.jp/global/download/pdf/Western_Blot_Protocols.pdf
Article CBB staining protocol with higher sensitivity and mass spect...
Article Total Protein Analysis as a Reliable Loading Control for Qua...