I am looking for antibody against human cell membrane which will circularly stain the cell membrane only. Could anyone name such antibody? I want to avoid F-actin, microtubulin bcz they stain all over the cells.
Secondly, can I use antibody for WB for IF? My understading is, Yes. Could anyone share their opinion or experience?
Hi Abdullah,
I do not know a marker that stains all human cells. You could chose a marker that fits the celltype you want to stain.(like CD45 for lymphocytes)
In the datasheet of your antibody it should be stated whether your Ab is suitable for both WB and IF.
In WB you stain a denatured protein; in IF you stain a naturally folded protein. Depending on the epitope recognized by your Ab is will work on IF.
First check the datasheet and otherwise just give it a try.
With kind regards,
Ronald
Hi Abdullah,
if you only want to stain the membrane (e.g. for size determination or cell boundary visualization) you can also use a cell membrane dye like this one: https://www.thermofisher.com/order/catalog/product/C37608?SID=srch-srp-C37608
Just be aware that they are not compatible with permeabilization, which you would need for IF. The other strategy, as mentioned, will be the IF of suitable receptors (depending on the cell type).
Good luck
Chris
Hello Mr. Al Tarique,
there is no such a thing. Instead of using antibody you can try dyes (like CellMask which is shown by Christoph Paone).
Good day!
Hi Chris, Thanks for your suggestion. Could you please mention one or two membrane receptors.
Hello
Yes , what kind of cells do you want to stain ?
and kindly see this link
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4709840/
Good Luck
Hi Abdullah,
I can recommend you Connexin43 (Cx43)ABs for cardiomyocytes and Fibroblast Surface Antigen (FSA) ABs.
Both have worked perfectly in my practice.
Cheers
Good morning Abdullah,
Regarding the first question, you could find a list of some markers for human macrophages here:
https://www.bdbiosciences.com/documents/cd_marker_handbook.pdf
Article The Identification of Markers of Macrophage Differentiation ...
About the second question, the possibility to use antibodies for WB for IF is really epitope-dependent, as with WB you are staining a denaturated protein whereas with IF you are staining a folded protein. However, all the info should be reported on the datasheet, and it is not so uncommon to find antibodies suitable for both purposes.
Good luck!
For mouse macrophages i used CD11b antibody from a hybridoma cell line from DSHB which was purified in our lab (http://dshb.biology.uiowa.edu/M1-70-15-11-5-2_2). See also our article in journal of innate immunity https://www.karger.com/Article/Pdf/442944
For human macrophages this antibody should work but is not 100% specific so you could select another one from the list that Daniele posted here (e.g. CD14). If it works in WB I never tested, but regarding the datasheet there is little hope...
kind regards
For surface staining human macrophages I usually use CD11b antibody for FACS analysis.
I think some company provide antibody usable for FACS and same time WB (e.g. Santa cruz).
Best of luck
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