Anti-CD62P antibody [AK-6] (ab6632)

No signal in western blots of human platelet lysates compared to previous lot.

ANSWER:

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I have issued a free of charge replacement.

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I wish you the best of luck with your research.

Where on the protein does this antibody bind?

ANSWER:

Thank you for your enquiry regarding ab6632.

Unfortunately this antibody has not been epitope mapped so I cannot tell you exactly which amino acids it recognises. However this clone, AK-6, was studied as part of the Adhesion Structures section of Leucocyte Typing V, Volume 2. In this report the epitope for this clone is localised to the SCR domain of CD62P.

I hope this information helps. Please do not hesitate to contact us if you need any more advice or information.

I have been using ab6632(Mouse monoclonal to P-Selectin) for microscopy and had good results as I mentioned in my last email. Your support told me I could use thia Ab for WB in either non-reducing or reducing conditions. I have tried non reducing conditions, using the Ab at 1:1000 and 1:250 with both secondary to HRP and AP with no results. I am currently doing a reducing western. I would like to know what specific part of the P-Selectin antigen does this Ab bind to? Secondly, does this cross react with E-Selectin?

ANSWER:

Thank you for your enquiry.

This antibody is specific for CD62P. To our knowledge it has not been tested for cross-reactivity with E-selectin (CD62E). As discussed in the reference given below, this antibody was raised against a human platelet glycoprotein which was confirmed to be platelet GMP-140 (or CD62P). The exact epitope recognised by this antibody has yet to be determined.

Skinner, M.P. et al. (1989). Characterisation of human platelet GMP-140 as a heparin-binding protein. Biochem Biophys Res Commun. 164(3):1373-1376.

We would usually recommend a concentration of 1-10ug/ml for the use of antibodies in Western blotting. Based on this estimate you may need to increase the concentration of antibody used further (i.e. to 1:100).

We hope that this information is of assistance.

Will CD62P work on other species? On formalin fixed tissue?

ANSWER:

This antibody has been shown to cross-react with human and Rhesus monkey. Other species have not been tested. This antibody is suitable for use on formalin fixed tissue.