Anti-Corticotropin Releasing Factor Receptor 2 antibody (ab75168)

This customer has highly interested in ab75168 (Anti-Corticotropin Releasing Factor Receptor 2 antibody), she would likes to order this antibody and test in IHC-Fr with rat sample, would you please help this customer to create a discount code, the information of this customer as follow:
Could you please help this customer to solve the problem? Thanks for your kindly help.
Best regards

ANSWER:

DISCOUNT CODE: XXXXXXXX
Expiration date: 13 Sept 2012

I am very pleased to hear you would like to accept our offer and test ab75168 in [Rat]. This code will give you: 1 free [PRIMARY ANTIBODY/PROTEIN] OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for [Rat] and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

human, WB, HeLa as positive control; Will the abs cross-react with each others targets?

ANSWER:

Ich habe mich mit dem Labor in Verbindung gesetzt, und HeLa sind eine geeignete Positivkontrolle auch im untransfiziertem Zustand.

Des weiteren sollten die beiden Antikörper nicht mit dem jeweiligen anderen Protein kreuzreagieren.

What is the chance that this antibody will cross react with Corticotropin Releasing Factor Receptor 1?

ANSWER:

This antibody will not react with Corticotropin Releasing Factor Receptor 1.


I hope this is helpful. Please contact us again if you have any further questions.

Hello,   I purchased antibodies against Corticotropin releasing factor receptors 1 and 2 (ab59023 and ab75168). I used both antibodies on western blots of cell lysates from Wild type, CRF1 KO, and CRF2 KO bone marrow derived mast cells.  There appears to be non-specific binding of both antibodies to both CRF receptor KO cells i.e. a band is present when anti-CRF1 antibody is used with CRF1 KO cells.  Can these antibodies bind both receptors?  What region of the receptor do they bind?  The receptors are fairly similar.  Do you have any suggestions for optimizing the blot?  I followed the protocols provided by the people who reviewed the products on the website.  

ANSWER:

Thank you for your reply. I am sorry that I misunderstood your original question. I have contacted the lab about the possibility of CRF1 and CRF2 cross-reacting. The CRF12 antibody cannot recognize CRF1. For the CRF1 antibody, the immunizing peptide KLYYDNEKCWFGKR used to generate and affinity purify CRF1 does not match entirely CRF2. There is sufficient difference with the comparable region in CRF2 to expect no or poor cross-reactivity. If you believe that the antibodies are cross reacting with each other, I would be grateful if you could send me some of the data that you have generated so that I can have a look at it and also pass it along to the lab to get their opinion. I hope that we can resolve this issue for you and that you can continue with your experiments.

Hello,   I purchased antibodies against Corticotropin releasing factor receptors 1 and 2 (ab59023 and ab75168). I used both antibodies on western blots of cell lysates from Wild type, CRF1 KO, and CRF2 KO bone marrow derived mast cells.  There appears to be non-specific binding of both antibodies to both CRF receptor KO cells i.e. a band is present when anti-CRF1 antibody is used with CRF1 KO cells.  Can these antibodies bind both receptors?  What region of the receptor do they bind?  The receptors are fairly similar.  Do you have any suggestions for optimizing the blot?  I followed the protocols provided by the people who reviewed the products on the website.

ANSWER:

I am sorry that you are having difficulties using some of our antibodies in western blot. From the data on the website, it does appear that in addition to recognizing the bands of interest, for ab59023 a 50kDa band and for ab75168, a band at 48kDa, both antibodies do give non-specific bands at various molecular weights. In your western blots, are you seeing a band at the right size and the non-specific bands or just the non-specific bands? For ab59023, the epitope region is between ******of the human sequence and for ab75168 the epitope sequence is ******of the human sequence. I would be happy to help with some protocol suggestions, could you provide me with the protocol that you using, blocking conditions, antibody concentration and incubations times etc. I will then see if I can make any recommendations that would help.

I look forward to your reply and helping you resolve this issue.