What is the best loading control for Western Blots for serum exosomes?

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What is the best loading control for Western Blots for serum exosomes?

To compare expression differences of a certain protein in serum exosomes which are derived from differnent disease state, what is the best loading control for Western Blots for serum exosomes?

08 September 2019 9,003 2 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

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Anyone that can help with details in phosphopeptides enrichment?

Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...

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Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

Why does intensity drops as concentration increases?

Is there any reason for decrease in fluorescence emission intensity of HSA,BSA when increase the concentration ?

28 February 2021 3,653 4 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

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Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

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What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

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How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Jana Deitersen

Hello Wenjuan,

Very generally speaking, ideally you use total protein stain (e.g. REVERT™ Total Protein Stain for the LI-COR System). For exosomal markers you can try CD63, CD81, CD9, TSG101 or ALIX.

See https://www.ncbi.nlm.nih.gov/pubmed/31035355 or Article Proteomic comparison defines novel markers to characterize h...

Jacó Joaquim Mattos

Hi!

I agree with Jana Deitersen .

Alternative methods for you.

There is a total protein normalization procedure with ponceau staining. Look for the question in research gate: Can Ponceau staining of membranes be used as a loading control ?

Also, if you have money and the equipaments, try stain free normalization. Article: Stain Free Total Protein Staining is a Superior Loading Control to β-Actin for Western Blots.

Good luck!

Jacó