Peptide labeling efficiency?

Hi all, I am currently trying to label my peptides by reductive dimethylation. I was just wondering how to evaluate the labeling efficiency ? One of the way that I think of is to compare the peak...

06 July 2018 8,838 1 View

What would be the best technique to get nuclear extract for a transcription factor study?

Hello you all, I am trying to isolate nuclear extract to study the transcription factors. I am mainly working with cell lines, I have found two kits for this....

05 June 2018 4,996 7 View

Jurkat T-cells culture for cytotoxic drug-treatment?

Hello, I am planning to enrich O-GlucNAc proteins in Jurkat T-cells by metabolic labeling before performing drug-treatment with Cisplatin. I have some practical questions regarding this: 1. Is it...

09 October 2017 7,157 0 View

What is Mascot 5 version in Scaffolds analysis?

I have tried to identify and enrich phospho-peptides by using TiO2 beads (control vs cisplatin treated Jurkat T-cells). Then I searched all the raw files using Proteome Discorverer 1.4 against...

08 September 2015 9,805 4 View

Do you have any advice on the isolation of BCL-2 protein family from Jurkat T-cell lysate?

Dear research gate members, I am trying to detect all the members of the BCL-2 protein family in T-cell extract. However, normal procedure in proteomics experiments did not work efficiently....

11 December 2014 9,019 4 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

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

How to increase the lower detection limit of ELISA?

Hi all, I am trying to establish an ELISA method for the quantification of a recombinant protein injected into mice. In the meantime I am trying to make calibration curves with the purified...

27 January 2021 8,702 3 View