Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210)

Inquiry: Hello, P16 INK4 antibody (ab54210) conc is not mentioned in the web site. It is only written as 100 ul but you are recommending 1ug per millions cells for sort/FACS. Please let me know 1 ug means how many ul. Thanks.

ANSWER:

Thank you for contacting us. The concentration for this product is not determined as it is from ascites. Concentration values are only meaning for purified antibodies, since those from ascites may contain proteins other than the protein of interest. Therefore we recommend you use the recommended dilution for your application.


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

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Can you please provide a recommendation for performing heat mediated antigen retrieval on free floating sections?

ANSWER:

Thank you for your phone call. Please find the reference I mentioned attached. I hope this helps, please let me know if you need any other information or assistance.

Incorrect banding pattern in western blotting.

ANSWER:

Thank you for contacting Abcam.
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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.
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The credit note ID is for your reference only and does not automatically guarantee the credit.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

I want to order your p 16 ink4a antibody.
So, I like to know exactly thatimmunogen of p16is Ni-NTA purified recombinant P16 expressed in E. Coli strain BL21 (DE3).
What'smean and what's your sequence of p16 aminoacid?
That is very important to me.

One more thing, p16 is working at rat liver? You used that with positive control.

Let me know if you have convenient time.

ANSWER:

Thank you for contacting Abcam.

Unfortunately for this antibody, the expression vector was provided for us for protein expression from another sourceto be used as the antigen. We were never supplied with the protein sequence for it as that information is proprietary, so we actually do not know the immunogen sequence for this particular antibody, and the immunogen is not available for commercial purchase.
I can confirm however that the antibody will react with rat liver.
My apologies we cannot be of more help with this, but if there is anything else I can help you with, please let me know.

Will ab54210 work in ICC-IF. Has this been tested?
For what reason were HeLa cells chosen to test in flow cytometry? We are not sure they express this protein. Can HeLa really be used as a positive control?

ANSWER:

Thank you for your telephone enquiry yesterday afternoon.

As discussed on the phone, I am sorry to confirm that to our knowledge, ab54210 has not been testedin ICC-IF. All testedand guaranteedapplications and species are listed on the online datasheets, and these are updated as soon as we receive any new information.

I can offer a discount off a future purchase if you buy ab54210 now, test it in ICC-IF and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab54210 in ICC-IF. I will then send a discount code. This code must be issued before purchasing ab54210 so please wait for my reply before ordering.

2. Purchase ab54210 either by phone, fax, or online (www.abcam.com).

3. Test it in ICC-IF.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: http://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab54210 turns out to be unsuitable for ICC-IF, you will still receive the discount on your next purchase after your Abreview has been submitted.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Regarding the positive control, I can confirm that HeLa cells would be a suitable positive control. these have been used successfully in flow cytometry with this antibody. I also notice that many other CDKN2A/p16INK4a available from Abcam and other suppliers are tested in HeLa.

There are many publications which confirm the expression of CDKN2A/p16INK4ain HeLa cells, for example:

p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary Cells
Kimberly M. McDermott et al
PLOS Biology
http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.0040051

I hope this will be helpful to you. Please let me know if you have any questions about the testing discount offer and I would be happy to help you further.