Rac1 and Q61L Rac1?

03 April 2017 3,597 4 View

Protein Sample Purity for Single particle cryo EM?

Hi everyone, I am using a chemical cross-linker to form a complex of three proteins (A-200kDa; B-21kDa; C- 17kDa) for Cryo EM. I get around 20-30% cross linked complex and the size of this complex...

03 April 2017 5,330 6 View

Arginine effects on crystallization?

Hi everyone, Can Arginine inhibit the crystallization process of a protein? I use 500mM Arginine to suppress the aggregation of my protein (30mM HEPES,pH7.5; 150mM NaCl; 5% Glycerol; 500mM...

06 July 2016 855 6 View

Suggestions on Getting crystals of protein complex in the presence of additives?

05 June 2016 6,853 4 View

Suggestions on purification of proteins having membrane binding regions?

Hi everyone, I am trying to express and purify a protein in BL21 (DE3) and arctic express RIL cells, which has a membrane binding region. I am using Triton X100 and CHAPS as detergents for its...

04 May 2016 5,081 5 View

Any suggestions on how to break protein aggregation?

what chemicals can be added during purification to inhibit protein protein interaction?

01 February 2016 1,562 15 View

Purification of proteins with Basic PI

Has anyone expressed and purified proteins with basic PI? I am purifying a protein which has a PI of 8.78. I see a band on SDS-PAGE when protein and solubility tag are together. However, After...

01 February 2016 9,868 8 View

Any suggestions on a problem of homogeneous aggregation during protein purification?

I am expressing a recombinant human protein with MBP solubility tag in E.coli BL21 cells. I am getting a very good expression, However my Gel fiteration profile shows aggregation of cleaved as...

10 November 2015 9,458 8 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

How do you prepare a porous crosslinked Polyvinyl alcohol and Glutaraldehyde membrane or film?

I am trying to prepare a Polyvinyl alcohol membrane for carbondioxide adsorption

08 February 2021 7,493 3 View

Is it possible to quantify recombinant protein in the supernatant using SDS PAGE ?

I would like to quantify a recombinant protein (with native signal sequence) secreted by Pichia pastoris using the SDS PAGE. Since my standard protein (plant origin) is not pure, I would like to...

07 February 2021 3,880 9 View

How to measure the concentration of a labeled protein with no tryptophan or tyrosines?

Hey I'm trying to measure the concentration of a protein I've purified and then labeled with Atto 647 flourescent dye. The protein has no tryptophans or tyrosines (although does have...

04 February 2021 4,482 4 View

How to prepare protein samples in EMSA assay?

Recently l was studying EMSA assay, but l didn't find any detailed information about protein sample. we can use two kinds of protein, including crude protein extracts and recombinant proteins. but...

02 February 2021 10,047 3 View

Difference between assessing EC50 in presence of physiological levels of human serum albumin and alpha1 acid glycoprotein vs. % human serum?

01 February 2021 1,825 1 View

Alpha synuclein lack structurw (i.e some of the helix part goes into turn ), any cause behind this?

I am trying to simulate alpha synuclein (1xq8.pdb) in GROMACS using GROMOS53a6 ff. I have run it for 200 ns. At the end of simulation alpha helix protein losses its helical structure. I am not...

31 January 2021 4,285 3 View