Detection of phosphorylated transmembrane proteins with WB?

Aleksandra Petrovskaya @Aleksandra-Petrovskaya

06 September 2023 0 7K Report

I need to detect phosphorylated IGF1R in cell lysate and brain lysate with Western Blot. Are there any life hacks for detection of such proteins?

Badges
Science topic

More Aleksandra Petrovskaya's questions See All

HPLC baseline problem. Baseline problem?

Hello, I noticed a problem with my HPLC analysis and wanted to ask for help. During my analysis I've noticed repetitive problems with the baseline: it is dropping and then (usually) going back...

20 May 2024 6,501 3 View

What’s are microorganisms adaptation to live in high sugar concentration (osmophile microorganisms)?

Hello I am looking for information about osmophilie microorganisms and specifically their enzymes. If there are any industrially important enzymes from microorganisms living in environment where...

19 May 2024 136 1 View

How to do root sampling for transcriptome analysis?

I am preparing an experiment in which I would like to sample roots of sunflowers grown in pots (substrate is soil) in later stages of the plant development. Can anyone tell me, what is the best...

29 April 2024 8,775 1 View

What's the specificity/tolerance of TaqMan probes for differences in sequence?

Hi, Does my TaqMan probe recognize the gene only if the sequence has 100% coverage with the probe or will I also get signal if there is a SNP or even a few bp difference? And if a few bp are ok...

09 April 2024 6,856 3 View

Why ions affect enzyme activity different in differ buffer?

I have tested enzyme activity (b-galactosidase) with cation ions in different buffer and the results differ dramatically. For example, while I used MES buffer calcium acts as an activator but when...

11 February 2024 3,135 2 View

Cell lines from Accegen - quality?

Hello, Does anyone have any kind of experience with ordering cell lines from Accegen? I'm wondering if they are trustworthy and about the quality of provided cell lines?

26 November 2023 3,757 1 View

Do you know of any tourism journals that have a mentoring programme for beginning reviewers?

I think that mentoring programmes support in meeting publishers' expectations.

05 October 2023 2,583 1 View

How do you get information about the number of chromosomes or DNA content?

I need to get information about the number of chromosomes or DNA content. I have seedlings from in vitro that were frozen at -20 degrees after liquidation of the culture. They are seedlings from...

28 September 2023 6,373 2 View

Would you like to attend our Summer School on Energy and Society?

https://webmagazine.unitn.it/en/evento/sociologia/115480/esa-rn12-environment-and-society-summer-school

10 March 2023 3,806 3 View

How many times should the permeability of the transwell-membrane covered by endothelial cells differ from the uncovered membrane?

To model the BBB using transwell-membrane, I use bEnd.3 cells. The permeability of the monolayer is assessed by bypass of the fluorescent tracer Na-fluorescein through the monolayer.

06 March 2023 6,105 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 503 3 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 966 3 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

Consequences of excess bases in the MCS after digestion?

Good morning, I am trying to produce a recombinant protein. Unfortunately I one of the two restriction enzymes that I use do not cut at the end of the MCS. After the digestion 15 nucleotides of...

17 July 2024 2,787 4 View

Phosphorylation sites prediction?

Hello, Does anybody have idea about any software or database that can give information about the possible phospho sites located in a protein? I am trying PhosIDP but its not working or loading...

15 July 2024 1,940 3 View

How to detect phosphorylation on Western Blots?

Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?

15 July 2024 2,055 3 View

How can CHARMM-GUI be used for protein-ligand interaction ?

My protein is a transmembrane protein, therefore i need to construct a transmembrane using CHARMM-GUI. Can someone please suggest me a tutorial for the same.

24 June 2024 5,044 1 View