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Questions related to Protein Purification Techniques
I am working on beta lactamase. I have constructed a clone containing beta lactamase gene, transformed and expressed protein using Penicillin in E. coli. Now need to isolate protein. I have tried...
12 December 2015 1,046 7 View
in order to check the efficiency of AKTA
03 December 2015 5,658 7 View
I expressed protein and unfortunately it became insoluble and remained in cell pellet. How do I get protein from pellet, so later I can use it for antibodies production?
26 November 2015 9,661 13 View
I am using MMP-2 to cleave the GPLG-VRGK short peptide. I have tried it in water? Does MMP-2 is active in water? Could you please tell me why it is not active in water? Which will be the best...
23 November 2015 373 12 View
I want to precipitate an aim protein binding to nuclear acid, so i add a final concentration of streptomycin sulfate to 0.83%. But the precipitation didn't have my aim protein. Should I increase...
11 November 2015 7,864 3 View
Membrane filters to concentrate 10Kda protein
11 November 2015 794 3 View
Hii everyone. I have a problem in case of pGEM T vector. I ligate my gene of interest and tag together into pGEM T vector but after transformation i get no colonies. I followed this protocol for...
11 November 2015 2,575 1 View
The undersigned is in the process of purification of immunogenic proteins from the cancer tissue. Total protein available with us is partially purified. The immunogenic band is resolved at the...
11 November 2015 7,805 0 View
Hello everyone, we are using High Capacity NeutrAvidin™ Agarose beads to separate biotinylated proteins. For screening experiments we'd like to reuse the beads. Therefore, I am wondering if anyone...
11 November 2015 628 1 View
Hi! I have been extracting histones using 0.8M HCl. I have used acid-soluble fraction for SDS-PAGE and subsequent WB. I found that I can actually stain extract for other nuclear proteins - for...
11 November 2015 2,137 1 View
Could anyone give me some suggestions about the "45Ca2+ blot overlay assay "? I searched for the procedure but there are some points that I could not understand. The general protocol is...
11 November 2015 1,217 4 View
Dear all, I am currently designing a lysis buffer for my protein of interest, expressed in E. coli BL21(DE3).The protein's solubility crucially depends on the correct formation of its single...
11 November 2015 2,014 3 View
By immunohistochemistry, we are getting an increased expression of various proteins but the mRNA expression of the genes coding for these proteins is showing an opposite pattern. Can anyone...
11 November 2015 2,165 1 View
I am studying a protein involved in DNA damage pathway. For my experiments, I wish to treat HEK293 cells with genotoxic agents: bleomycin and florouracil and UV. My protein of interest is...
11 November 2015 7,306 0 View
Hi I'm trying to purify an over-expressed membrane protein which I suspect is stuck in inclusion bodies. I extract the proteins using 8M Urea in a 50mM Tris-HCl (pH 7) buffer containing 20mM...
11 November 2015 4,135 7 View
Is anyone out here who has done vector purification with TFF? I'm looking to compare experiences with systems, especially anything above 1 liter. What did you use, what were your experiences,...
11 November 2015 8,477 1 View
11 November 2015 4,212 13 View
How can we distinguish between Primer dimer and non specific amplification using gel electrophoresis and melt curve?
11 November 2015 1,720 3 View
I am attempting in situ PCR with paramecium. Has anyone tried in situ PCR in a similar organism with a pellicle? I trying to determine the parameters for fixation and protease treatment. Any help...
11 November 2015 3,020 1 View
Can we trust the 260/280 ratio? Do you have an alternative method to propose? Thank you. Nt
11 November 2015 7,686 4 View
Hello everyone,As a novice in this field, I would like to obtain a bioactive refolded protein from inclusion bodies. My his-tag protein is 77kDa cloned in pET32a (+), pI 5.84 and contain 8...
11 November 2015 424 14 View
I am looking for a tag that allows to purify recombinant proteins in mammalian cell with high affinity and high specificity in non-denatured condition. Can someone advise me what it is the best...
11 November 2015 6,066 6 View
Could someone suggest a protocol for protein label, i'm using a protocol on which i wash the protein with ethylacetate-ethanol after Precipitation of protein with 20% TCA, i have a doubt that i...
11 November 2015 5,786 1 View
If I want to purify prion protein scrapie (PrpSc) from brain tissue of mouse and in the first step I need to put protease inhibitor cocktail in PBS, then homogenize brain in it. I have 100x...
11 November 2015 4,875 6 View