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I would like to request information on one of your antibodies, Anti-CLLD8/SETDB2 antibody (ab5517). I was wondering if this antibody works for mouse SETDB2 |
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ANSWER: |
Thank you for your inquiry. I BLASTed the immunogen sequence of ab5517 against mouse and it only has 55% homology. We generally say that anything with a greater than 85% homology would be expected to cross-react, so this is quite low. This antibody probably wouldn't work with mouse samples. However, we do have a goat polyclonal anti-CLLD8/SETDB2 antibody that is predicted to cross-react with mouse, ab13712. This immunogen is 100% homologous with mouse. However, since we have not tested ab13712 in mouse, we would not be able to guarantee that it would work. http://www.abcam.com/CLLD8-SETDB2-antibody-ab13712.html But, I can offer a discount off a future purchase if you buy ab13712 now, test it in mouse 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 ab13712 in mouse. I will then send a discount code. This code must be issued before purchasing ab13712 so please wait for my reply before ordering. 2. Purchase ab13712 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse. 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 ab13712 turns out to be unsuitable for mouse, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount. |
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BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 72248 DESCRIPTION OF THE PROBLEM Also purchased ab13712. Multiple bands with each antibody. Molecular weight of bands different for both antibodies. Not sure if I'm picking up the correct band and if so with which antibody SAMPLE Human cell lysates A549, jurkat, C8166, Daudi, HUT-78 PRIMARY ANTIBODY ab5517 or ab13712 SECONDARY ANTIBODY cell signalling anti-Rabbit-HRP #7074 abcam rabbit anti-goat6741-1 DETECTION METHOD chemiglow POSITIVE AND NEGATIVE CONTROLS USED none available SAMPLE PREPARATION RIPA + protease inhibitors, heated 5min TRANSFER AND BLOCKING CONDITIONS NuPAGE blotting buffer + 20% MeOH, 2hr. Blocked 5% milk in PBS-tween HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No WHAT STEPS HAVE YOU ALTERED? primary antibody concentration
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ANSWER: |
I'm very sorry to hear you are having problems with ab5517 and ab13712. You do not mention adding loading buffer to your samples prior to boiling, please make sure you add loading buffer containing SDS, DTT, beta-mercaptoethanol and bromphenol blue. Please also make sure you keep samples on ice until boiling and that protease inhibitors are added freshly. You do not mention how long you block the membrane for (and what temperature), I recommend trying 5%BSA in TBST for 1 hr, then rinsing a few seconds in TBST and incubating both primary and secondary antibodies in TBST. The dilution recommended for ab5517 is 1:5000 and for ab13712 is ug/ml but you may need to dilute more, I hope these suggestions help, please do not hesitate to contact us if you need further assistance,
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