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 10,004 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 2,937 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 7,564 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 4,314 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 1,350 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 1,238 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 2,862 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 9,239 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 6,244 0 View

Signal peptides in secretory proteins?

I wish to find out if all secreted proteins have a signal peptide. Also, if yes, are these signal peptides always cut?

07 August 2017 9,895 2 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Could you please suggests a tutorial or paper or any other materials to learn multiple gene expression in TCGA and GTEx databases?

I would like to analyze multiple gene expressions in normal and cancer tissue by using the data in the TCGA and GTEx databases. Please suggest a tutorial or paper showing step by step analysis of...

23 February 2021 9,007 3 View

Are there any databases for gene co-expression or expression pattern clustering?

I am currently working on gene clustering based on co-expression pattern in mouse brain. The problem is I do not have some solid way to test my result. Are there any suggestions for databases...

22 February 2021 4,374 3 View

Which CpGs have a significant effect on gene expression?

It is known that methylated CpG around TSS decreases gene expression. However, It is difficult to know exactly which CpG has a greater impact on gene expression. Are there any databases known...

20 February 2021 5,634 3 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

Purification of proteins from baculovirus-infected insect cells?

18 February 2021 6,665 6 View

Found that there was a multiple band when amplifiying targeted gene ?

i few days ago amplified specific gene using PCR with specific primers, but there was a multiple band in electrophoregram i found. out of 10 sample amplified, there was multiple band and there was...

18 February 2021 5,131 5 View