Phospho - PI3KD

July 09, 2015

Medicine, Division of Allergy/Immunology
Northwestern University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biorbyt

Product Name:

PI3KD (phospho - Tyr524) antibody

Catalog Number:

orb6720

I work on trying to understand the signaling mechanism of a surface receptor expressed on eosinophils and PI3KD is a potential signaling molecule. I selected this product because there are not a lot of these antibodies out there.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Peripheral blood human eosinophils

Primary Incubation

1:500 in blocking buffer (Rockland - MB - 070), shaking at 4*C O/N

Blocking Agent

Fluorescent Western Blotting (Rockland - MB - 070)

Secondary Incubation

1.5 : 10,000 in TBS-T (IRDye 680 from LI-Core)

Tertiary Incubation

NA

Detection

Odyssey imager

Results Summary

Eosinophil protein was isolated following treatment with stimulus or control.

Additional Notes

Do not sonicate the protein lysate, it removes any phosphorylation events and the Ab no longer works.

Related Categories

Image Gallery

Summary

The Good

When it works, the bands look great, but that is not always the case.

The Bad

It needs a lot of trouble-shooting and even then it's difficult to guess whether its going to work or not

The Bottom Line

It's an okay product, especially as it's the only one out there. You just have to be patient and keep troubleshooting.

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