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Anti-Phosphotyrosine antibody (ab9319)

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3 questions for ab9319

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Question 1

Wednesday 08-February-2006

I need a non-mouse anti-Phosphotyrosine antibody to use in immunocytochemistry assays on fruit fly tissues, labelling the plasma membrane. Your site does not mention ICC for this product, does that mean it does not work?

ANSWER:

  

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested for application in ICC. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image).

Please contact us again if you have any additional questions.

Question 2

Wednesday 17-September-2003

For the Immunoprecipitation you recomend acetone precipitation of the protein extract fallowed by SDS denaturation. Do you have a protocol for that?

ANSWER:

  

1. Add cold acetone:lysate sample, v/v=9/1. eg: 100 uL smaple add 900 uL acetone. 2.Centrifuge at 5000 rpm for 5 min 3. Reconstitute with the precipitate with same volume of PBS then add 0.5% of SDS to the solution with 10% SDS stcok. 4. boil for 5 min then cool down to room temperature before IP

Question 3

Monday 25-November-2002

As a positive control, you suggest to treat cell culture with vanadate for 30 min. Do you have a protocoll (concentration?) or reference for this kind of treatment? Many thanks

ANSWER:

  

You could use the mouse fibroblast cells as positive control.

First, remove the old medium and culture the cells in a serum-free medium for 24 hours. Then empty the dish again and treat the cell with the fetal calf serum media containing 1 mM Vanadium for 30 min. After that harvest the cells and use the cell lysate as positive control.

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