Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)

I am interested to buy Anti-Histone H3 (acetyl K9) antibody to use it in my ChIP experiment. but the thing is you have 3 H3K9 recommended for ChIP. 1- ab4441 2- ab10812 3- ab113674 Which one do you recommend is the best one for ChIP?

ANSWER:

Thank you for contacting us.

Antibody ab113674 is the newest antibody of the three and is therefore less thoroughly characterized. Antibodies ab4441 and ab10812 have both been published many times and have received positive customer reviews for ChIP, among other applications. Ab4441 is the more popular of the two, most likely because it has many more references.

Please do not hesitate to contact us if you need any more advice or information.

Please see customers technical inquiry below:



“We would like to know if the antibodyhttp://www.abcam.com/Histone-H3-acetyl-K9-antibody-ChIP-Grade-ab10812-support.html will work in paraformaldehyde fixed brains (rat), set in agar or tissue tek (cut by vibratome or cryostat, respectively). We only ask this question as the tech details describe that this antibody is good for paraffin fixed brains. It would still be appropriate to work in our set up?”



Looking forward for your response.





Kind regards,

ANSWER:

Thank you and your customer very much for your interest in ab10812.
To our knowledge, ab10812 has not been tested in IHC-Fr yet ,but is very likely to work in IHC-Fr since it is tested fro IHC-P. Therefore, I can offer a discount off a future purchase if your customer buys ab10812 now, tests it in IHC-Fr and submits 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 customer's feedback. The discount would be to the value of 1 Free primary antibody.
If your customer is interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know that your customer (please include their e-mail address) would like to proceed and test ab10812 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing ab10812 so please wait for my reply before ordering.
2. Purchase ab10812 either by phone, fax, or online (www.abcam.com).
3. Test it in IHC-Fr.
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 customer's 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 ab10812 turns out to be unsuitable for IHC-Fr, your customer will still receive the discount on their next purchase after your Abreview has been submitted.
Please let me know if you or your customer 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.

Dear Ladies and Gentlemen,
In the future (mid of the year) I want to perform ChIP on Chip experiments. For this purpose I´m planing to use
antibodies from your company which will be verified first in simple ChIP experiments. Our experiences are that the
efficiency of different lots of the same antibody largely varies from lot to lot in ChIP experiments, meaning that verifying the
ChIP grade of an antibody now to perform ChIP on Chip in half a year is meaningless if the lot of the antibody changes in near future.
Thus my question is, is it possible to order antibodies of a certain lot, verify their functionality in my
experimental set up and re-order the antibody with the same lot number in e.g.3-6 months?

ANSWER:

Thank you for your enquiry.

Unfortunately we are not able to "save" specific vials of antibody for customer that may wish to purchase them in the future. If you'd like I can recommend specific lots that have the most stock, which may give you the best chance to buy an antibody at a later date from that same stock. The downside with this scenario is that it will only increase the likelihood the specific lot will be available but not guarantee it.

Also, perhaps shortening the time frame for testing the antibodies might also give us a better chance to provide you with the lots you need.

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

Histone H3 (acetyl K9) antibody (ab10812): Sample: Chicken Cell lysate - nuclear (HD3) Loading amount:: 20ug

Application: Western blot

Blocking step:: Milk as blocking agent for 1 hour  · Concentration: 5%

Conditions:: Reduced

Incubation time:: 12 hours 

Detection method:: ECL

Exposure:: 1 hour, 0 minutes and 0 seconds

Bands:: Specific: 16 kDa Dilution: 1/100

Rating: Poor detection- I would look for a better antibody

Secondary antibody: Name:: Non-Abcam one was used: goat antirabbit immunoglobulins

Conjugation: HRP

Dilution:: 1/10000

ANSWER:

Thank you for your enquiry.

I am sorry to hear that you have been having difficulties with this antibody. I have read your review and I would like to make the following recommendations.

You are performing an approach that I would recommend for the detection of a cytosolic protein. However, as this antibody is targeting a modification of a single residue on histone H3 I would recommend that you increase the mass of protein that you are loading to 50ug. Furthermore to improve the results that you have been obtaining I would like to recommend that you perform a histone extraction of your chicken cells in order to enrich for the antigen. This antibody is tested in-house using purified calf thymus histones using an antibody dilution of 1:500.

We also employ BSA as a blocking agent in our lab testing. This often improves the signal and specificity and is something that I would recommend. For a detailed protocol please see the following link;

http://www.abcam.com/assets/pdf/protocols/Histone%20western%20blotting.pdf

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.