Standard RIPA buffer including protease and phosphatase inhibitors and 0.1% SDS diluted in 4X Laemmli Sample Buffer [62.5 mM Tris HCl, 1% LDS, 10% Glycerol, 0.005% bromophenol blue, pH 6.8] BME has not been added.
This lysate is prepared as a positive control for separation by SDS-PAGE and subsequent Western blot analysis. Lysates are prepared in denaturing buffer WITHOUT reducing agents (i.e. 2-mercaptoethanol (BME) or dithiothreitol (DTT)). If reducing conditions are desired, add a reducing agent prior to heating. Heat lysate to 95 degrees C for 5 minutes and rapidly cool. The recommended loading amount is 0.01-0.03 mg per lane.
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FAQs for HepG2 Lysate (NBP1-42569). (Showing 1 - 2 of 2 FAQ).
How was this lysate prepared? Please advise as I am trying to use this material as a control lysate for a kit I purchased.
This lysate was prepared using standard RIPA buffer with 0.1% Triton X-100, and 1% SDS and sodium deoxycholate. We used mechanical separation to remove the cells from the plate and also use a cocktail of protease and phosphatase inhibitors in the lysate buffer.
Good morning, we have to work with short-HepG2 Whole Cell Lysate 1.0mg to do adsorb on nanoparticles. We are chemical-physical. I would like to know if the sample handling under the hood (not flow alminare) will have a biohazard and what precautions should be taken.
NBP1-42569 is a whole cell lysate with the cells in a RIPA buffer. These cells are lysed and not longer viable. They are not considered a biohazard and no special precautions need to be taken.