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k.g.'s Content

There have been 2 items by k.g. (Search limited from 15-April 20)

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#145983 hellp! Need instruction manual for Omnifuge rt 3950

Posted by k.g. on 28 November 2012 - 10:37 AM in Lab Equipments

We just recently got a used Heraeus Sepatech Omnifuge rt 3950 bench top refrigerated centrifuge unfortunately it did not come with a user manual on how to operate the machine. I have a manual about the rotors and tubes but not about how to operate the machine. I have done a general internet search which came up with people selling the machine with the manual but not the user manual itself. I was wondering if anyone had the manual and would be willing to upload a pdf of some of manual or also know a good place where I could get a user manual for this machine?

thanks so much!

#133790 pro-collagen extraction from Mouse Tissues

Posted by k.g. on 30 April 2012 - 08:54 AM in Protein and Proteomics

Hi, new to the site. I am currently trying to extract procollagen from mouse organs, kidney and liver mostly.

I originally used a buffer with extraction buffer of
-20mM Tric-HCl
-150mM NaCl,
-1mM EDTA,
-1% Triton X-100.
On top of that I add 40ul Protease inhibitor cocktail (sigma) to every 1ml of extract buffer right before I use the buffer for extraction. This is a buffer that I used in the past and have success extracted other proteins with it but when I tried it to extract pro-collagen I got really poor results. I read the samples in an ELISA and had to use 10mg/ml total protein to get any reaction, which was, still very week and did not make it onto the curve. I can typically use 5mg/ml of total protein for the other proteins that I extract which I are in far smaller quantities in the cell than collagen and procollagen. I know that collagen is insoluble and this is probably why I did not recover very much procollagen.

Does any body have any experience with extracting procollagen so that it can be run in an ELISA or how to extract Collagen to be run in an ELISA?


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