n this step, wash the cells with ice-cold PBS. Drain the PBS, then add ice-cold lysis buffer. Scrape adherent cells off the dish using a cold plastic cell scraper, then gently transfer the cell suspension into a pre-cooled microfuge tube. Maintain constant agitation for 30 minutes at 4°C. Centrifuge in a microcentrifuge at 4°C. You may have to vary the centrifugation force and time depending on the cell type. Gently remove the tubes from the centrifuge and place on ice, aspirate the supernatant and place in a fresh tube kept on ice, and discard the pellet. Q3: To make sure that you used a similar amount of samples, what important step should be done before proceeding the electrophoresis stage? Once you have determined the concentration of each sample, you can freeze them at -20°C or -80°C for later use or prepare for immunoprecipitation or for loading onto a gel. Q4. Why is it necessary to store the prepared lysates in a very low temperature? PLEASE ANSWER Q3 AND Q4
In this step, wash the cells with ice-cold PBS. Drain the PBS, then add ice-cold lysis buffer.
Scrape adherent cells off the dish using a cold plastic cell scraper, then gently transfer the cell suspension into a pre-cooled microfuge tube. Maintain constant agitation for 30 minutes at 4°C. Centrifuge in a microcentrifuge at 4°C. You may have to vary the centrifugation force and time depending on the cell type. Gently remove the tubes from the centrifuge and place on ice, aspirate the supernatant and place in a fresh tube kept on ice, and discard the pellet.
Q3: To make sure that you used a similar amount of samples, what important step should be done before proceeding the electrophoresis stage?
Once you have determined the concentration of each sample, you can freeze them at -20°C or -80°C for later use or prepare for immunoprecipitation or for loading onto a gel.
Q4. Why is it necessary to store the prepared lysates in a very low temperature?
PLEASE ANSWER Q3 AND Q4
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