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Protein: Detection, Quantification and Kinetics
Short Description: Protein overexpression checked using SDS-PAGE, quantified using Bradford's assay, confirmed by western blotting and activity determined using enzyme kinetics.
Type: Hands-on
Duration: One day (9.00 AM to 5.30 PM)
Fees: INR 2596 (INR 2200 + 18% GST)
Category (for payment): MBY Workshop Tier 2
Aimed at: College students (B.Sc. and M.Sc.)
Application Process: Click here for the details
Registration form (This link will be different for each workshop)
Application process (in brief)
Application process (in brief)
Check the above table to figure out whether suitable dates are available or not.
Note down the category of the workshop.
Head to SB Collect to pay the fees. Save the Receipt and the SB Collect Reference number. See here for details.
Fill the above Registration form using the Receipt and the SB Collect Reference number. Choose the workshop date. You will get an email acknowledgement from us. Registration is NOT yet complete.
We verify your payment and send Registration Confirmation Email. Your registration is now complete.
We strongly encourage the applicants to look at the detailed application process before paying the fees and filling the form.
Course Content
Course Content
1. Basic laboratory practices
1. Basic laboratory practices
Basic safety, handling of micropipettes and safe disposal of laboratory wastes. To know more about how to handle micropipettes, watch this video or practice using this simulation.
2. Understanding the technique of protein overexpression and extraction
2. Understanding the technique of protein overexpression and extraction
In the interest of time, extracted protein will be provided. To know more about protein isolation, watch this video.
3.Total protein estimation by spectrophotometry
3.Total protein estimation by spectrophotometry
After the cell lysis, the amount of total protein is quantified by spectrophotometry which detects the concentration of the final reaction product. To know more about spectrophotometry, you can read this article or watch this video for how a spectrophotometer works.
4. SDS-Poly Acrylamide Gel Electrophoresis
4. SDS-Poly Acrylamide Gel Electrophoresis
The overexpressed protein is checked by SDS-PAGE to examine the protein expression profile of the samples and separate the proteins in a way similar to agarose gel separation of DNA. To know more about SDS-PAGE, please read this article or watch this video.
5. Transfer of protein and detection using an antibody (Western blotting)
5. Transfer of protein and detection using an antibody (Western blotting)
Proteins are transferred from the polyacrylamide gel to a membrane and probed with an antibody specific to the protein of interest. To know more , read this article or watch this video.
6. Enzyme assay to detect activity of protein
6. Enzyme assay to detect activity of protein
The enzyme assay is used to determine the amount of protein activity associated with each sample. The assays are performed in triplicates to gain confidence in the values measured. Enzyme kinetics, read this article or watch this video.
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