Anti-Bmi1 antibody [1.T.21] - ChIP Grade (ab14389)

Dear Abcam Tech support,   I’ve been working on mouse anti-human BMI-1 antibody (ab14389) on feline tissue.    IHC showed nice nuclear staining and we ran some Western blotting to get further evidence of cross-reactivity of this ab against feline tissue.   As you can see in the attached pic, we saw an intense band at around 37kDa (which we wanted to see).  However, we saw other bands at around 100kDa.  Interestingly, we saw similar MW band in the canine MCT cell line too.    Without the primary ab, we didn’t see any bands.   Of course we are curious about the higher MW bands, but we first want to get further support to say that the band around 37KDa is a specific one.   I’m wondering if you have a blocking peptide that can specifically bind to this ab (ab14389). Or you could give me some different advice, we’d appreciate a lot.   I look forward to hearing from you…

ANSWER:

I have just heard back from the lab and unfortunately the blocking peptide is not available for ab14389. Please let me know how boiling the samples for longer works out.

Abcam Tech support,  I’ve been working on mouse anti-human BMI-1 antibody (ab14389) on feline tissue.    IHC showed nice nuclear staining and we ran some Western blotting to get further evidence of cross-reactivity of this ab against feline tissue.  As you can see in the attached pic, we saw an intense band at around 37kDa (which we wanted to see).  However, we saw other bands at around 100kDa.  Interestingly, we saw similar MW band in the canine MCT cell line too.    Without the primary ab, we didn’t see any bands.   Of course we are curious about the higher MW bands, but we first want to get further support to say that the band around 37KDa is a specific one.   I’m wondering if you have a blocking peptide that can specifically bind to this ab (ab14389). Or you could give me some different advice, we’d appreciate a lot.  

ANSWER:

Thank for contacting Abcam. I have been in contact with the lab about what the band at ~100kDa is, as it seems that several Bmi1 antibodies recognize this high molecular weight band. Unfortunately they have not been able to identify what the high molecular weight band could be. It is possible that the protein could be in a complex with other proteins or even itself, by boiling for 10 minutes rather than 5 minutes, it could be that these complexes will get broken up and therefore you would not see the high molecular weight band. I am still waiting to hear back about our ability to provide the blocking peptide and will let you know when I more information. In the meantime if there is anything else I can help you with please let me know.  

I’m just wondering if this code below is active and I can use to get 1 free primary antibody?

And, if I use the code to purchase ab18976 () to use against feline tissues (IHC), can I get another code if I submit abreview?

Thank you

ANSWER:

Thank you for your enquiry.

The code provided is active if you have submitted your Abreview regarding your experience using ab14389 to detect feline protein. I see that you have in fact submitted your review so yes, the code is active.

For this promotion, it is required that one antibody, at the discretion of a Scientific Support representative, be purchased at the list price to receive a discount code. So you could not get another code by submitting a review for the free antibody. You can of course submit an Abreview based on your experience with the free antibody and you will receive points for that review. These points can be used for discounts on future purchases or Amazon giftcards.

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

I tried to create an account to submit a review but your website says that my email address has been already used.   I honestly don’t know what’s going on but that’d be great if you could check if there is my account already created (in that case, please send me some info to log on so that I can write a review).

ANSWER:

We have created this account for you. Whenever someone contacts us we need to make an account for the customer in order to log all emails back and forth.

Your account password is xxxxx. You can change it once you log on.

Please let me know if you have any other questions.

Yes, I’m interested in ab14389.

ANSWER:

I am very pleased to hear you would like to accept our offer and test ab14389 in feline. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for feline and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. 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.