Anti-KDM6B / JMJD3 antibody - ChIP Grade (ab85392)

Since we talked last time, I've been working really hard on KDM6B in mice. I tried Ab84200, it gives me inconsistant results. And I can't re-use the diluted antibody since I can't detectanything with re-used AB. I can't write a reviewbecause I am not even sure whether there is a band or not.

As for Ab85392, no matter what I try, noKDM6B can be detected with this antibody. I am pretty sure about it. On the datasheet, the band shown is detected in HepG2 cells which is a human cell line. Do you have solid data to support your claim that this ab can detect mouse samples?

I have run out both of the Antibodies. It only has about 40 ul. I had to buy another aliquote of Ab84200 which cost me $378.00. I think that is way over charge for something we are not even sure will work.

ANSWER:

Thank you for your reply.
I am sorry to hear that you are struggeling so much with these antibodies.

I have checked again the datasheet for ab85392: Mouse was shown to work in the publication that is listed on the datasheet:

Miller SA et al. Jmjd3 and UTX Play a Demethylase-Independent Role in Chromatin Remodeling to Regulate T-Box Family Member-Dependent Gene Expression. Mol Cell 40:594-605 (2010). WB, ChIP; Mouse. PubMed: 21095589

It might be of help to check this paper, or even to contact the authors for more experimental details if necessary. Also, the homology between human and mouse is 93%, and the antibody was raised against a mouse seuqence.

If you submit your inconclusive data for ab84200 (which has not been tested in mouse and is therefore not guaranteed), your testing discount code will become active and you can use it on another antibody purchase. But you wouldneed to submit your data (even if there is no band). The review has also a free text box where you can add your observations.

We do have a new lot for ab85392 in stock which you could obtain using the testing discount code. This is the only KDM6B / JMJD3 antibody which has been tested for mouse samples.

As for ab84200, since mouse is untested for this antibody, we do not guarantee it and therefore, I had sent you the discount code for trying it (according to our testing discount program). You just needed to submit your data/Abreview to receive a free antibody for it.

In terms of troubleshooting advice, it might be worthwhile to includea positive control in your experiment (e.g. HepG2 cells or a mouse derived cell line (e.g. NIH 3T3)).

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

Catalog number(s): 85392
Product(s): Rabbit polyclonal to KDM6B / JMJD3 - ChIP Grade
Survey results
------------------------------------------------------------------------------------------Telephone section
Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
A. Yes
Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
A. Yes

Protocol advice section
Q. Did Abcam's Scientific Support give you protocol advice?
A.Yes
Q. How did you feel about being given protocol advice?
A.I was not expecting to be given protocol advice, but was happy to receive this
Q. Did the product work successfully after following the advice?
A.No
Q. Why have you not tried the protocol advice? (Select all that apply.)
A.
Free of charge replacement section
Q. Did Abcam's Scientific Support give you a free of charge replacement?
A. No

Q. Did the replacement product work successfully?
A.
Credit note / refund section
Q. Did Abcam's Scientific Support give you a credit note or a refund?
A.No

Q. Would you have preferred a different outcome from receiving a credit note or refund?
A.
Overall satisfaction section
Q. How satisfied are you with how the complaint was handled?
A.Neither satisfied nor unsatisfied
Q. Could we have done anything to better resolve the problem?
A.I have so much trouble with KDM6B antibodies from Abcam. None of them succeed in western as claimed by datasheet. And those antibodies are ridiculously expensive. You can try to develop better antibodies and charge less.
Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
A.No, I would not mind
Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.
A.Very good company. Just some of the products don't work as the way your company claimed.

ANSWER:

Thank you for your participation in Abcam's post complaint survey.

I am sorry to hear that the KDM6B antibodies do not work for you satisfactorily.

According to our correspondence, your are working on mouse samples and only ab85392 has been tested so far in mouse. This antibody gave you bands at the incorrect band size. Since none of the other 2 KDM6B antibodies were tested in mouse, I had sent you ab84200 together with a testing discount code for trying the antibody in mouse.

In order to redeem the discount code, you would need to submit an Abreview with your negative data. Then you can receive a free antibody on a future order from us. I would like to encourage you to submit the Abreview.

Please let me know if there is anything else that I can do for you.

mouse kidney samples
sees non-specific band at 40 kDa, but no band at expected MW of 180 kDa
loaded protein from 10 to 300 ug
transfer OK
Ab: 2 ug/ml overnight 4 degree
block: milk
secondary: OK

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab84200.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is notcovered by our Abpromise guarantee since mouse is not yet tested with this antibody ab84200. However, I am sending you a tetsing discount code in that regard. Please see below for more details and the actual code.

I wish you the best of luck with your research.

====== TESTING DISCOUNT CODE ======

DISCOUNT CODE: xxxx
Expiration date: xxxx

I am very pleased to hear you would like to accept our offer and test ab84200 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouseand 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: http://www.abcam.com/collaborationdiscount.

I know ChIP antibodies are special in respect of reacting with antigen after fixation sample. What does your ChIP-grade mean?

ANSWER:

Thank you for your reply.

When an antibody is characterized as ChIP-grade, this means that the product is validated for use in ChIP and is guaranteed to work in this application.  If for some reason the product did not work well in ChIP, you would be eligilble for a replacement or credit under our Abpromise guarantee.

I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.

I am wondering that I am going to use ab85392 for ChIP assay (Genome-wide), but there is one concern. You are showing a nice exact size band (180kDa) in WB, but another size band (44kDa) is seen.

Is this antibody available and feasible for specific ChIP experiment? I am afraid that non-specific 44kDa could also react with Jmjd3 antigen.

ANSWER:

Thank you for contacting Abcam regarding ab85392.

I have examined the datasheet for this antibody and can confirm that this antibody has been validated for ChIP.  Unfortunately, we do not know the identity of the 44kDa band.  Validation in ChIP was based on a publication in which a customer used this antibody in ChIP.  The reference is:

Miller SA et al. Jmjd3 and UTX Play a Demethylase-Independent Role in Chromatin Remodeling to Regulate T-Box Family Member-Dependent Gene Expression. Mol Cell 40:594-605 (2010). WB, ChIP; Mouse. PubMed: 21095589

This antibody is guaranteed to work in ChIP and we are happy to replace or credit it per our Abpromise. 

I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.