Is it necessary to acetone precipitate plasma proteins before iTRAQ labelling for mass spec?

More Dr. Narendra Kumar Sharma's questions See All

Anyone can help with endnote style for shock?

I tried to search output style for shock journal but didnt find. If any one want to share style or information about same style reference of other journal.

10 November 2015 5,120 1 View

Anyone have idea how to find a miRNA target site?

I wants to find miRNA target site by manual or tool available foe access

08 September 2014 1,803 16 View

Can anyone suggest a reliable tool for the design of qRT-PCR primers when used with sybr green?

Can any body suggest or share idea about primer design for qRT-PCR for gene Perilipin 2 step by step for beginner?

04 May 2014 5,498 10 View

How do you make a ROC curve from tabulated data in R?

I want to make an ROC curve from tabulated data using R. Can anyone share the codes or any tutorial for doing this?

02 March 2014 7,223 3 View

Is it possible to identify different oxidized or reduced state of a protein in mixture by mass spectroscopy?

I want to study a signaling molecule which shows reduced or oxidized nature.

01 February 2014 7,013 4 View

Can anyone want to share method for PCA plotting or suggest a free tool for PCA analysis of proteomic data?

Thanks in advance.

01 February 2014 2,749 4 View

Which methods is best for comparing the Plasma proteome between two groups by LC-MS?

I have plasma samples and want to compare two groups to identify differentially expressed proteins by LC-MS. There are several approaches like label free and iTRAQ. I want to know which method is...

01 February 2014 3,401 11 View

How can we inhibit a particular signaling pathway?

To silence genes of a particular pathway to study the significance of said pathway.

31 December 2013 9,811 9 View

Is there any free software to make hierarchical clustering of proteins and heat maps with expression patterns?

31 December 2013 1,381 11 View

Is a cut-off filter suitable to exchange buffer from protein samples?

I experienced, that when I used 3 or 5 kd cut-off filter tubes to exchange buffer for proteomics work, the flow through have some protein content. Theoretically it should not have happened. In...

31 December 2013 7,753 3 View

Can you connect an HPLC to a Mass Spec only at a certain time point?

Can anyone explain this method? Especially the last statement where it says only at 1.5 to 2.5mins was the MS/MS connected to the UPLC. How is that possible, is it a feature in this specific...

11 August 2024 8,141 3 View

Baseline drift in HPLC? What causes this?

Hello, Why do i see this baseline drift when i compare my blank (black) to the sample (blue)? Any suggestions as to why this happened? Thank you!

11 August 2024 3,770 4 View

GC-MS retention index prediticon?

Hello experts, Does anyone know any free software about retention index prediction ?

08 August 2024 7,403 2 View

How to calculate CCS for Sodiated adduct ions and Multiply Charged Ions?

I'm currently working on calculating the collision cross section (CCS) for various ions, and I'm facing challenges when dealing with sodiated and multiply charged ions. Most of the resources I’ve...

08 August 2024 8,329 0 View

Where to find a gene list for CRISPRa/i library screening of regulatory factors that affect pathogenic Th17 differentiation in PBMC?

I want to perform a CRISPRa/i library screening for candidate regulatory factors that affect pathogenic Th17 differentiation in PBMC. But I am wondering how to define the size of gene library, so...

31 July 2024 2,150 0 View

Hello dear researchers. I would like to know what is a normalized mass in an EDX analysis?

EDX analisys

31 July 2024 5,947 2 View

Gas chromatography RT detection?

I am working on algal extract to which gas chromatography (Not GC-MS) spectrum I want to discover. My question is can we identify specific compounds using retention time if I compared the RT with...

29 July 2024 8,034 4 View

Can the limit of quantification (LOQ) of an analytical method fall outside its linear dynamic range, or must it always be within it?

Can an analytical method's limit of quantification (LOQ) be outside its linear dynamic range, or is it always required to be within it? Please provide a thorough explanation supported by verified...

29 July 2024 7,198 9 View

Where to start the Analytical model for Dry sliding wear of metal surface ?

I am doing research on wear mechanism of the additive manufactured metal parts under dry condition. Please share the reference to start the analytical modelling of the wear prediction.

29 July 2024 6,115 0 View

MAGeCK Plotting Error: Logarithmic Axis Must Have Positive Limits?

I have been running a MAGeCK test command on the terminal for a CRISPR screen to rank sgRNAs and genes based on the read count tables. However, I get only the plot of the top-ranked genes...

28 July 2024 9,811 1 View

Joseph V Moxon

Hi Narendra

I've not used iTRAQ on plasma samples, however, was advised to avoid precipitating any protein sample due to inherent problems with resolubilising in an iTRAQ compatible buffer. We recently got by this problem by using centrifugal filters to buffer exchange extracted proteins in 0.5M TEAB prior to any processing. From there we performed Bradford estimations and continued with the labelling protocol as per manufacturer's advice.

Dr. Narendra Kumar Sharma

Thanks Joseph Moxon for suggestion

Can you share the plasma sample preparation methods in detail with me.

Not used plasma - the analysis I performed was using crude tissue extracts, however I found the attached paper useful in terms of iTRAQ processing. The paper detailing my experiment (including the buffer exchange) has just been accepted - will forward a copy to you once I get the proofs if you think they'll help.

Joe