How can I perform a serine protease phenotypical detection?

Can i use the protease inhibitors during cell membrane vesicle preparation ？?

I am currently working on a project that involves extracting cell membranes, for which we disrupt the cells using sonication. During the initial extraction process, we add protease inhibitors to...

30 July 2024 7,077 1 View

Are the apoptotic cells is positive for γH2AX ?

It has been documented that apoptotic cells themselves can induce phosphorylation of serine 139 on H2AX (γH2AX) due to DNA fragmentation during apoptosis (doi: 10.1074/jbc.275.13.9390). As γH2AX...

28 July 2024 7,983 2 View

Can it is possible to find the cleaved sequence when a protein cleaved by a heamaglutanin protease (HA/P) by any bioinformatics tools?

Bioinformatics tools like peptide cutter

15 July 2024 6,453 1 View

How do you determine the Ki of an inhibitor for an enzyme which also undergoes substrate inhibition?

There are three factors involved in the activity, the substrate acting as a substrate, the substrate acting as an inhibitor and another inhibitor for which you want to measure the Ki.

08 July 2024 4,988 1 View

How to incease the yiald of recombinant protein expression ?

Hello esteemed scientists, I am endeavoring to increase the yield of my recombinant protein. It is a 120 kDa protein tagged with MBP and features three intrinsically disordered regions with pI...

12 June 2024 9,616 8 View

Can you explain the potential toxicity and overdosemanagement of commonly used antiplatelet drugs?

Antiplatelet Therapy: Mechanisms, Indications, Administration Can you explain the potential toxicity and overdosemanagement of commonly used antiplatelet drugs?

05 June 2024 6,447 1 View

How to prepare 0.28 U/ml working solution of Acetylcholinesterase inhibition assay for anti-Alzheimer activity?

I have purchased Acetylcholiesterase from Electrophus electricus (electric eel), C3389-2KU, following details are written on the enzyme vial. Type-VI S lyophilized powder 200-1000 Units/mg...

31 May 2024 6,444 1 View

Can anyone interpret why the absorbance in alpha amylase assay get increased with increased standard (inhibitor) concentration?

Can anyone interpret why the absorbance (DNS assay method) in alpha amylase assay get increased with increased standard (inhibitor) concentration? it should be decreased with increased standard or...

26 May 2024 4,753 2 View

Does the catalytic activity of a lyase, such as a protease, change upon the formation of a μ-gelzyme?

21 May 2024 9,403 0 View

Should the target miRNA level decrease after the miRNA inhibitor transfection when checked with qPCR??

I have a question regarding target miRNA expression after transfection with antisense miRNA (miRNA inhibitor). I understand that the expression of the target miRNA will increase after mimic...

20 May 2024 5,920 4 View

Steingrimur Stefansson

Hi Morteza, the easiest way to detect proteinase activity in these bugs is zymography

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC97462/

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC128331/

Morteza Rabi Nezhad Mousavi

Hi. may you explain more!!! because i dont know zymography . I need method for detection but these articles that you introduced are simple and dont explain efficiently. thanks steingrimur

Hi Morteza, here are some papers on the zymography technique

http://docsdrive.com/pdfs/scienceinternational/sciintl/2013/70-75.pdf

http://www.ncbi.nlm.nih.gov/pubmed/9193704

A list of papers on serine proteinase zymography

http://scholar.google.com/scholar?q=serine+zymography+&btnG=&hl=en&as_sdt=20000005&sciodt=1%2C21&cites=8902318142592596246&scipsc=1

thank's a lot

Chitra Latka

Did zymography work for you ?