Rabbit Control IgG - ChIP Grade (ab46540)

Hello,

I'm using Rabbit Control IgG - ChIP Grade (ab46540)
for my chip experiment.
Do you have equivalent for rat and mouse? I don't find them,


Thanks in advance,

ANSWER:

Thank you for your inquiry.

Indeed we do have also polyclonal control fractions of mouse or rat IgGs.

For the mouse IgG, we do have the IgG fraction ab37355 http://www.abcam.com/index.html?datasheet=37355 (or use the following: http://www.abcam.com/index.html?datasheet=37355).

For the rat IgG, we do have the IgG fraction ab37361 http://www.abcam.com/index.html?datasheet=37361 (or use the following: http://www.abcam.com/index.html?datasheet=37361).

These two products have been used as controls so far only in ELISA and flow cytometry to our knowledge. As they have however no known specificity, I would expect them to work also in ChIP. If you would like to test them in ChIP, please do let me know, as I might can provide you with a testing offer.

Otherwise, if you are looking for a monoclonal rat or mouse antibody as a control, we do have also monoclonal isotype controls. Voici la liste de nos anticorps isotypes controls: http://www.abcam.com/index.html?pageconfig=catalog_byclass&intClassID=718&strStartChar=A,B,C,D,E,F,G,H,I,J,K,L,M

Please do not hesitate to contact us again if you have anyquestion, if wecould help you in the further selection of an antibody or if you are interested in testing the above mentioned antibodies in ChIP.

Compare the ab46540 to the Anti-GFP antibody (ab290), which one you suggest
if I use it as negative control for both H3K4me3 antiobody and H3K9me3?

ANSWER:

Thank you for your reply.


The isotype control Rabbit Control IgG - ChIP Grade ab46540 can be used as a negative control for any rabbit IgG unconjugated antibody. It does not show species specificity, meaning it's just matched to the species of the primary antibody you're controlling for not the species of the sample you're testing (mouse or human).

This isotype control can be used with ab8898, the Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade and ab8580, the Rabbit polyclonal to Histone H3 (tri methyl K4) - ChIP Grade. This is probably the best way to go instead of using a non-histone related ab like GFP.


Unfortunately we do not sell formaldehyde so this is something you will have to purchase separately.


I hope this information helps. Please contact us with any other questions.

dear correspondent,

We have lately purchased several Abs from Abcam and i have a question related to the purification method.
In the datasheets of several monoclonal antibodies is mentioned that they are purified as 'IgG fraction'. Could you exlain to me what that means? Is a anti-IgG column used or an protein A/G column (or any other technique)? This is very important information to us, as we need to consider protein A/G contamination to explain data obtained in our R&D dept. in experiments using these antibodies.

More specifically, the question relates to the following antibodies (data sheets of all those abs mention 'IgG fraction' in 'purity' annotation):

ab3609-100 (anti-Aurora B)
ab51841-100 (anti-TATA binding protein)
ab37373 (goat control IgG)
ab46540 (rabbit control IgG)


Thanks in advance and hope to hear your answer soon,

ANSWER:

Thank you for your enquiry.

I have confirmed the purification details of the antibodies you listed and have updated the datasheets where necessary:

ab3609 (Aurora B)
Protein G purified

ab51841 (TATA binding protein)
Protein G purified

ab37373 (goat control IgG)
Proprietary combination of ammonium sulfate precipitation, ion exchange, and/or other chromatography techniques.

ab46540 (rabbit control IgG)
Affinity purified against the antigen.

I hope this will be helpful. If you require any further information on the purification of other products, or any other questions, please do not hesitate to contact me.

I need some help designing my ChIP experiment.

ANSWER:

Thank you for contacting us.


First, I'd like to point out some resources on our website which may help:


http://www.abcam.com/index.html?rid=310


A Beginner's Guide to ChIP - Abcam


As we discussed over the phone, you may want to start with our ChIP kit (ab500)since this has everything you need for the reaction, including the positive control antibody ab1791.


http://www.abcam.com/ChIP-Kit-ab500.html


In addition, you'll need the two antibodies you were interested in, and their matching negative controls.


Anti-Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)


http://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-ChIP-Grade-ab8580.html


Rabbit Control IgG - ChIP Grade (ab46540)


http://www.abcam.com/Rabbit-Control-IgG-ChIP-Grade-ab46540.html


Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)


http://www.abcam.com/Histone-H3-tri-methyl-K27-antibody-mAbcam-6002-ChIP-Grade-ab6002.html


Mouse IgG3, kappa monoclonal [MG3-35] - Isotype control (ab18394). This one isn't tested in ChIP, but I don't see a reason why it wouldn't work.


http://www.abcam.com/Mouse-IgG3-kappa-monoclonal-MG3-35-Isotype-control-ab18394.html


You'll also need the housekeeping primers, like GAPDH, ab83588 and ab83589.

http://www.abcam.com/hsgapdh-chip-primer-exon-1-forward-ab83588.html

http://www.abcam.com/hsgapdh-chip-primer-exon-1-reverse-ab83589.html


That should be enough to get you started, but take a look at the resources mentioned above and let me know if you have any additional questions.

Dear, We lately bought the rabbit control IgG antibody. We would like to rebuy it but need further information on the following question. Is this antibody purified via protein A chromatography or not? This is important information for us if we want to assess the quality of the ab. This is not clearly mentioned on the datasheet. If you dont know, please also indicate so, so that we can consider protein A contamination in the sample to explain data generated by us using this ab. Thank you very much,

ANSWER:

Thank you for your inquiry.
I can confim that the antibody was affinity-purified against the antigen itself (bound to agarose).
Protein A or G was not used.
I hope this information is helpful. please do not hesitate to contact me with any further questions.