i have been unable to get band of catalase Native PAGE. I am using 10% polyacrylamide gel. I follow the staining procedure of Woodbery et al., 1971 and also Thorup et al 1961. Please can someone explain the extraction procedure of sample and all that.
Hello Imteyaz,
you don't give enough details to help you more specifically with your question (e. g., sample type, origin, extraction method, etc.). Therefore I can only recommend to consult the paper in the attachment. Maybe you can find the answer to your question therein?
Regards, Christian
Article Measurement of superoxide dismutase, catalase and glutathion...
Hello,
I agree with the previous answer. In addition, I would suggest to pay extreme attention to the use of sonicators to prepare cell extracts. Many catalases, for example bacterial catalases, are readily inactivated during sonication, unless there is a very tigth control of temperature.
Hi Imtiaz,
please ensure the following things inorder to get bands,
1. the crude extract that has been isolated has to be fresh or atleast stored immediately at -80 degrees celsius. If you use freeze thawed sample it will not work.
2. Add DTT for mobility while loading the sample
3. Try a lower concentration of stacking and separating gel 5 and 7% respectively
I agree with the previous researcher. Be careful about protein concentración used and run the gels using cooling systems (or within a coold room....to avoid increase of temperatura during the electroforesis).
Good luck
Rosa María Martínez Espinosa
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