Signal peptides in secretory proteins?

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How do I purify rabbit IgG4?

I want to purify IgG4 antibodies from immunized rabbits. I have tried to look for protocols to no avail. Commercial kits on the other hand exist for humans and a few other animals but I have not...

03 April 2019 10,153 3 View

Any bioinformatics tools to predict immuno-modulatory or immuno-regulatory epitopes in proteins?

Hello everyone, we are characterizing some proteins in the context of vaccine development and we are interested in determining there are any epitopes that may rather lead to immune evasion. Don't...

03 April 2019 3,080 3 View

How do I avoid impurities from my SEC purification?

Hello everyone, so I expressed this protein and I am trying to further purify using SEC and still having the impurities in the fraction I expected to have the protein. How do I eliminate these. I...

03 April 2019 7,714 3 View

How do i prepare water saturated phenol:chloroform mixture (5:1)?

I am doing RNA, DNA and protein extraction from the same sample and my protocol recommends this water saturated phenol:chloroform mixture. I am not not very versed with how to go about with the...

03 April 2018 4,443 8 View

What are the best (recommended) tools for 3D protein structure prediction, refinement, and validation; also what is the best tool for MD simulation?

Hello everyone, I am trying to do some hands-on bioinformatics and will be grateful to get your orientation.

31 December 2017 1,479 5 View

How do I interpret protein 3D structure refinement data?

Hello everyone, I did 3D structure prediction (using I-TASSER) for a protein and submitted the best model for structure refinement (with GalaxyRefine). Unfortunately, I don't have a very good...

11 December 2017 1,362 0 View

What is the best buffer to us in antigen-capture ELISAs?

Hello everyone, I am about doing an antigen capture ELISA and a little confused about which buffer can be used in absorbing the capture antibody to the microplates. I am using Nunc MaxiSorb 96...

10 November 2017 2,991 3 View

What is the effects of coating ELISA plates with antigen for longer than recommended durations?

Hello everyone, So I planned this ELISA to do, unfortunately I coated plates and for some reason I am not able to continue with the experiment as planned. So basically, instead of coating...

09 October 2017 9,378 4 View

Autoinduction of pGEX5x3?

Hello everyone, I wish to express a gene cloned in pGEX5x3 through auto-induction and compare with the normal IPTG induction. Any one has experience doing autoinduction with pGEX vectors?

08 September 2017 6,429 0 View

Isolation of inclusion bodies?

I am working with a pET30a+ construct and my protein is C-terminal His-tagged. My protein is forming inclusion bodies and I need to get a protocol for isolation of a pure protein from this. An...

07 August 2017 2,097 4 View

How to increase simulation box size?

We intend to study the interaction between peptides and polymer (like PP, PE and PS) through MD simulations using Martini force fields ( Martini 2 for PP and Martini 3 for PE, PS). We have...

08 August 2024 4,842 0 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

Unusual intensity drop in some sections of chromatograms in DDA?

Hi, we have measured tryptic peptides using both DDA and DIA method on QExactive. In DDA replicates i saw unusual intensity drops occurring at the same sections of chromatograms in DDA replicates...

07 August 2024 3,218 4 View

How to analyze multiple phosphorilation sites?

Hello, I am currently analyzing some phosphoproteomics data, but I have peptides with multiple phosphorylation sites or phosphorylations together with carbamidomethylation or oxidation. How can I...

04 August 2024 8,432 3 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

Recovery rate after peptide desalting using Sep-pack C18 1 cc Vac Cartridge?

Recently I'm trying to use Sep-pack C18 1 cc Vac Cartridge (Waters: SKU: WAT054955) to desalt digested peptides. However, the recovery rate is usually 40-60%. What is the normal recovery rate?

31 July 2024 6,544 11 View

Problems with ion exchange chromatography?

I'm having problems with ion exchange chromatography. After applying my sample (a fluorescent protein), a preferential path inevitably forms. The matrix I use is DEAE-Sephacel. Could someone...

27 July 2024 6,605 0 View

How do regenerate Amberlite XAD 4 resin for further cycles of use for removing 0.5% Sarkosyl from recombinant protein solutions?

I use 0.5% sarkosyl to solubilise recombinant proteins contained in Inclusion bodies of E. coli. sarkosyl very effeciently solubilizes proteins from Inclusion bodies and I am trying to use...

25 July 2024 5,001 2 View

How do nutrient transporters and aquaporins interact under abiotic stress in plants?

"The Correlation between Nutrition and Transport Mechanism under Abiotic Stress in Plants: A Comprehensive Review" by Muhammad Saleem, Jianhua Zhang, Muhammad Qasim, Rashid Iqbal, and Li Song,...

23 July 2024 9,109 0 View

Is it possible to conjugate an NHS ester to the N-terminus of a peptide ON RESIN?

So i have a few peptides (synthesized by SPPS and not cleaved from resin) and I want to add a maleimide label. We have NHS-PEG-Maleimide and from literature it is evident that the free amine group...

18 July 2024 4,075 3 View