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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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protein extraction
Posted by: kum81 (IP Hidden, Junior member, 10)
Date: October 24, 2005 05:49AM
I want to know thw methodology for the extraction of extracellular protein of a bacteria. and their estimation and resolution on PAGE.
Re: protein extraction
Posted by: hazariv (IP Hidden, New member, 2)
Date: November 3, 2005 04:29AM
Hi,
If u r talking about secreted proteins, best way to extract is Ammonium sulphate precipitation at its iso-electric pH. The resultant precipitate can be pelleted down using a centrifuge and can be resolubulized in a suitable buffer or just water for further analysis. Ammonium sulphate can cause some problems in SDS PAGE so TCA precipitation can be helpful. To process it in higher volumes think about EBA etc, Find the reply useful or not- get back to me. regards
Re: protein extraction
Posted by: mamuller (IP Hidden, Junior member, 10)
Date: November 29, 2005 10:03AM
Protein separation is dependant of lengh of gel, maximum gel lengh provide maximum separation resolution. However typical lengh of mini gel is 8 cm, and resolution is sufficient. In most cases operator do not run the full 8cm lengh, as separation is sufficient after 6-7 cm running distance.
The best method for extracting the maximum of protein in gel is to use 1) To provide maximum protein recovery, Staining of protein MUST NOT have adhesive to protein in its formulation, only staining sb010 provide visual staining during process, no need to end staining process, using clean water remove staining. 2) GeBAflex-tube electro eluting protein from gel, minimum garantied recovery is 70%, visit www.geba.org for methode and process instructions. 3) have a search at www.biowww.net gebaflex-tube, you will find GeBAflex-tube protein extraction appreciation Contact me directly at miler-m@zahav.net.il for further details Maurice Maurice Muller
Bl21 proteases
Posted by: Samiaifa (IP Hidden, New member, 1)
Date: December 23, 2005 04:53AM
Has someone any idea about residual proteases recognition sites in BL21?
I am expressing a recombinat protein in BL21 and each time I detected it cleaved in the same site according to the same obtained sizes of 2-3 peptides. Thank you for your help Sami
Re: protein extraction
Posted by: kalyankrishna (IP Hidden, New member, 1)
Date: October 5, 2007 05:27AM
sir
send isolation of bacterial quorum sensing proteins related to intra, extra cellur proteins please ......................................... my id ; rambiosvu@gmail.com thank you sir
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