Why only few proteins identified from a total protein lysate run on a 1D gel subjected to LCMS analysis followed by Mascot search?

Dear all, I am wondering why i am getting only less proteins identified from a total protein lysate separated on a 1D gel. Did the in-gel digestion by cutting individual lanes in to 24 bands,...

04 May 2016 9,243 7 View

Why can't I see any protein spots after a 2D run?

10 November 2014 7,645 16 View

Does the foam formation during protein isolation have any impact on the protein quality?

During sonication/lysis after adding the lysis solution, protein extraction results in foam formation. Does this have any negative impact on protein quality? please provide me with suggestions Thanks,

10 November 2014 9,906 6 View

Why are proteins not fully soluble in the buffer after methanol-chloroform precipitation?

We precipitated the proteins using methanol-chloroform protocol and redissolved the obtained protein pellet in Tris/PBS buffers. But the proteins are not completely dissolved leaving behind a mass...

10 November 2014 3,857 4 View

Can anyone help with 2D gel images seems to be pixelated while cropping the images in PD Quest software?

We have taken the gel images of a 2D run and was trying to analyze those images in the PD Quest Basic software from Biorad. But the images are not clear when we are cropping it and seems like the...

07 August 2014 3,023 2 View

Why do I see a change in pressure when the HPLC instrument is in idle condition?

It seems like there is a frequent change in the pressure when the instrument is in idle condition, without running any samples having an isocratic flow. The column we are using is Poroshell-120,...

06 July 2014 2,635 8 View

What column and solvent composition should we have to use to do HPLC profiling of Vitamin K1 and K2?

We have bought Vitamin K1 and K2 standards from sigma and did the HPLC analysis of K1 by dissolving it in diethyl ether, but we didn't see a peak. We have used Agilent Poroshell-120, 150x2.1mm,...

05 June 2014 5,630 3 View

What buffers can we use to isolate proteins to perform 2D gel electrophoresis and what is the difference between a pharmalyte and an ampholyte?

I have seen a few papers mentioning like they have used urea, chaps, DTT and Pharmalyte for protein extraction. Is pharmalyte an ampholyte? What's the difference between a pharmalyte and an...

04 May 2014 3,695 5 View

What are the components of a lysis buffer for protein extraction to perfrom 2D gel electrophoresis?

What are the components or what lysis buffers can be used to extract proteins from cells so as to perform 2D gel electrophoresis?

04 May 2014 505 15 View

Which type of 96 well plate is suitable for suspension cells?

Since suspension cells have to be spun down before removing the media, I am just wondering weather there could be any advantage of using round bottom 96 well plates rather than flat-bottom 96 well...

02 March 2014 5,516 9 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

After a reference from Int J Body Compos Res, can any body help?

Hi, I am after the reference below, my library says it cannot obtain a copy either locally or internationally, any help appreciated! Chris Wang ZM, Heshka S, Wielopolski L, Pi-Sunyer FX, Pierson...

03 March 2021 6,193 1 View

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03 March 2021 5,360 2 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 View

How to compare different concentrations of nanoparticles for characterization from ICP-MS?

I took measurements of different concentrations of 15 nm and 5 nm Au nano particle solutions. I'm trying to determine which concentration I measured gives the best reading overall. I also...

02 March 2021 7,626 1 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Any research papers for extraction of nutrient deprivation on macroalgae? Analysed by HPLC-UV and GC/MS. Possibly from macroalgae genus Rhizoclonium?

I am doing my thesis on the nutrient deprivation on the macroalgae Rhizoclonium sp. I will be analysing the levels over a period of roughly 5 days by HPLC-UV and GC/MS. I will be using seawater as...

28 February 2021 6,477 1 View

Is there any published or unpublished statistics on dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, US, UK or any other country?

Are there any published or unpublished statistics or papers on the dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, USA, UK, or any other country where vaccination has started at...

28 February 2021 4,237 1 View

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View