Contact experts in Protein Purification Techniques to get answers
1,396 views 784 posts
Questions related to Protein Purification Techniques
I am preparing the TBST buffer for the western blot. The most common NaCl concentration is 150mM. However, I found tow the recipe from BIO-RAD with 500mM and 150mM NaCl concentration respectively....
19 June 2017 322 5 View
Hey guys, I'm having some difficulty purifying my 46 kD GST fusion protein, specifically, I'm having issues with proteolysis and purification. I appreciate all suggestions, however time and money...
06 June 2017 5,714 6 View
I am unable to get the protein at soluble fraction from plant but has managed to solubilized in bacteria using tag. Is there any way to find interaction partner of this protein using healthy plant...
06 June 2017 7,715 2 View
Hi all,I am trying to express a protein in E. Coli and I need to make sure 1) that the protein is in the soluble form and 2) that this protein is secreted. I am brand new to the bacterial world,...
06 June 2017 1,172 4 View
hi, i am trying to purify an 8kda protein.PI 6.95, HIS sumo tagged. after adding sumo protease to my elution sample plus 2mM DTT .DAILYSIS BUFFER being 20mM NaH2PO4 500mM NaCL..when i again bind...
19 May 2017 9,710 9 View
For purification of this protein i used lysis buffer( 20mM tri s HCl pH=8,with 0.5M Nacl and 0.2M imidazol), (20mM NaPB pH=6.5,with 0.5M Nacl and and 0.2M imidazol), (1x PBSl pH=7,and 0.5M...
05 May 2017 6,525 2 View
Greetings to all, I have been purifying a His-tagged membrane protein (ion channel to be precise) following a standard protocol and everything seems to be fine till Cobalt column purification...
05 May 2017 9,572 4 View
I'm currently working on the purification of EPO which is secreted from CHO cell . for final step of purification I have to applied IMAC-Fe(III) for an especial goal.(Endotoxin removal) could any...
04 April 2017 8,584 10 View
Hello, I would go for His-tag purification and I was wondering which company would be the best to order Ni-TA column? Do you have a preferred one or all of them would be the same? Thanks in...
04 April 2017 6,552 4 View
I tried 8M Urea buffer for plant protein extraction and nickel purification. I equilibrated the nickel beads with TEV cleavage buffer which contains 50mM tris, phosphate, 300mM NaCl. Then I added...
21 March 2017 5,391 6 View
I purified my proteins from plants in 7M urea buffer through nickel purification. Then I changed purified proteins into a native condition buffer for overnight (Tris and phosphate) for other...
03 March 2017 6,292 6 View
If I do a dialysis with the help of a dialysis membrane of my protein and then perform SDS-PAGE to check my desired band...... Excise it and purify... Can I do the sequencing of it??? As I am...
03 March 2017 6,512 4 View
I need to add ascorbic acid into solid medium. I prepared the medium, adjusted pH to 5.4 (pH for ND medium), autoclaved it, added ascorbic acid (0.58 mM; sterilized by syringe filter), and then...
03 March 2017 3,476 3 View
pI above 9
03 March 2017 544 4 View
I have a question regarding to your Amylose resin kit https://www.neb.com/products/e8021-amylose-resin I read in the protocol...
02 February 2017 9,626 7 View
The AcrA protein has purified from the E.Coli using a buffer containing Tris-Hcl, NaCl, PMSF, Triton-X and imidazole
02 February 2017 7,180 2 View
I need to purify serum glycoprotein present in very low concentration (2-10 ng/mL). Current method (sepharose bound monoclonal antibodies with subsequent elution on a filter plate) works, but...
01 January 2017 4,244 4 View
Does anyone know how to extract protein bands from Native gel to detect the product of the enzyme activity? I need the pellet from the band for further detection and to get rid of the gel.
01 January 2017 9,572 3 View
We are doing crystalization for a protein, which could exist as rod-like oligomer in cells. We expressed the protein in bacterial and in insect Sf9 cells. After purification procedures, protein...
01 January 2017 9,768 4 View
Solubilization buffer 2M Urea, 1mM DTT, 1mM EDTA & 10mM Tris.
01 January 2017 3,939 4 View
Hi, can any one suggest me, i am performing western blot to detect tnf alpha in brain tissue. i am using 12%gel and wet transfer method, tried with NC and PVDF. When i stain the gel i could do the...
12 December 2016 10,025 5 View
HIF1A protein gets stabilized after some time, but we are not able to see the band (stabilization) on western blot after various time points.
12 December 2016 8,510 6 View
hello,everyone. I need to solve this problem in western blot , i did the cell lysis by NP40 then protein assay kit to quantification the protein . i load same amount of protein 70 ug but...
12 December 2016 180 4 View
Hi! I'm trying to purify the main egg white proteins using an anion exchange chromatography column (Mono Q HR 5/5) in a FPLC system, but I have a lot of problems because the back pressure of the...
12 December 2016 8,351 9 View