Why do we use ammonium bicarbonate as a buffer salt in Ingel digestion for MS ?

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Which proteomics software is best for performing IBAQ analysis?

I have protein samples which I need to identify. I loaded it on SDS PAGE. Now each band may contain two or more proteins. Can I estimate relative abundance of those proteins through IBAQ? If yes,...

05 June 2019 6,131 3 View

Why are my gel pieces not getting destained for in-gel digestion?

My gel pieces are not getting destained. I have kept it for more than 6 hours now. I m using 50mM Ammonium bicarbonate and Acetonitrile in ratio of 7:3.

04 May 2018 1,828 1 View

How to properly store the sds gel pieces for in gel digestion?

I have already cut the gel pieces and would like to perform in gel digestion in future. So I had freeze them in 1% acetic acid. Can I still process those samples?

31 December 2017 3,059 1 View

Is proteomic data analysis in R (using RforProteomics) better than any other proteomic software?

My project involves analyzing proteome of my sample. I have more than 100 samples for analysis. Would proteomic analysis in R help me? Is it better than other softwares?

31 December 2017 6,581 8 View

Why do I get smear on SDS-PAGE?

I am working on snake venom proteins and wanted to resolve it on SDS PAGE. I have attached image of my gel. I am getting a smear. Is there any way to resolve it better?

04 May 2016 9,119 5 View

What is polymorphic marker?

I was reading an article where they have mentioned alleles of polymorphic markers. they are talking about U2 tandem arrays.

02 March 2016 8,035 2 View

How does DNA get compressed in stacking gel?

I know that it gets sandwiched between glycine and Cl ions But how does it get compress literally?

01 February 2016 7,311 2 View

After running SDS, why do we put our gel in solution of acetic acid and methanol?

I was reading SDS PAGE and methodology it was mentioned, I searched it online but i couldn't find the answer.

01 February 2016 506 5 View

Can anyone give me examples of gene-lists and gene-sets used in GSEA?

I wanted to understand gene set enrichment analysis. For that I need some practical examples of gene-lists and gene-set used in this technique

09 October 2015 7,558 4 View

How to do northern blotting to check expression of HSP proteins in frog ?

I have to quantify expression of HSP proteins in frog. So what could be best possible technique to do that?

04 May 2015 5,343 3 View

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 2 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 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

What is the best practice for resuspending cell pellets during competent cell preparation (both electro and comp)?

I'm fairly new to making homemade comp cells and have done so with some mild success. What is generally an acceptable way of resuspending comp cells during wash steps? I've been told to swirl the...

31 July 2024 8,309 0 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View