I am trying to quantify a protein in a glass tube, but most of it is adsorbed to the walls. Does anyone have methods / protocols to release the protein, and suggestions for the container to use in the detection test (will do a western blot I think, unless not sensitive enough)
Thank you!
Sarah
Hello, Sarah.
Add your lysis buffer (~300-500 uL depending on the tube size). Seal tightly not to leak. use horizontal roller mixer for blood tube.
Good luck.
Depending on your protein, glassware should be avoided altogether. Low-binding Eppendorf tubes (LoBind) might be a good option.
How do you quantify your protein?
If you load your sample on SDS PAGE fo western blot to avoid this glass sticking I advice to use low binding tubes or siliconized tubes. If you keep to use your glass tubes one coating with BSA faty acid 5% should reduce this sticking .
Hi, thank you all for your answers.
My problem is that I cannot change the container. It is a study for a final product, and the container should be a glass vial. I cannot add BSA or other agent in the glass initially because it would change the product. My question is really about desorption. How do you release a protein stuck to a glass wall?
- Badges
- Science method