Anti-acetyl Lysine antibody - ChIP Grade (ab21623)

Thanks for your suggestion

But I already ordered -competitor product- for our work (but I'm not sure it is best for me)
The products you recommendedare better than -competitor product-for our works?
If it is, I will order again..
My plan is to use dynabead (invitrogen) for IP


Best regards,

ANSWER:

Thank you for your reply.

Since the antibody you bought is not from our catalogue, I cannot comment of the suitability for your planned experiments.

If this antibody is tested and guaranteed (as we do with our antibodies) for IP and CHIP, then this antibody should most probablybe fine. But of course I will not be able to guarantee this and therefore suggest to get in contact with the company that produces this antibody and requestmore information on it.

I hope this information is helpful and wish you good luck for your experiments.

I want to selectively enrich lysine-acetylated peptides from tryptic digested protein mixutre

LC-MS/MS will be used for identification

Can you recommand some antibodies for me?

ANSWER:

Thank you for your inquiry.

I am assuming that before subjecting the samples to LC-MS/MS, the enrichments of the acetylated lysine containing peptide is planned to be done with immunoprecipitation?

I am happy to confirm that we offer two antibodies specific for acetylated lysine that are tested and guaranteed for IP (ab21623 is also CHIP tested and guaranteed).

Please follow these links to review the respective datasheets:

ab21623

http://www.abcam.com/index.html?datasheet=21623 (or use the following: http://www.abcam.com/index.html?datasheet=21623).

ab80178

http://www.abcam.com/index.html?datasheet=80178 (or use the following: http://www.abcam.com/index.html?datasheet=80178).

We also have extensive protocols for IP on our homepage:

http://www.abcam.com/index.html?pageconfig=resource&rid=11385

http://www.abcam.com/index.html?pageconfig=resource&rid=11382

I am not especially familiar with the requirements that samples have to adhere to for LC/MS/MS. But maybe elution of protein with little antibody contamination is preferable? Therefore I suggest the following protocol:

http://www.abcam.com/index.html?pageconfig=resource&rid=11415

I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

J’aimerai réaliser une Immunoprécipitation (IP) de peptides acétylés, suivi d’une analyse en protéomique (LC-MS/MS). Les peptides acétylés proviennent d’une digestion tryptique de protéines extraites de cellules.

Pouvez-vous alors m’envoyer un devis sur le produit suivant ou d’autres si vous pensez que d’autres produits seraient plus adaptés à mon étude.

rabbit anti-Ac-lysine: Abcam ref: ab21623

A quelle protéine puis-je le couplé A ou G avec sépharose. Merci d’ajouter le produit approprié au devis.

Avec 100µg de ce produit combine de peptide acétylé puis-je espérer capturer. Merci

ANSWER:

Merci de nous avoir contactés.

L'anticorps ab21623 est en effet un bon choix pour l'immunoprécipitation de peptides acétylés. Nous n'avons pasd'information supplémentaire sur le rendement de cet en anticorps en IP, cecidoitêtre déterminé par l'utilisateur.

ab21623 est un IgG de lapin, nous recommandons donc l'utilisation de billes couplées à laprotéine A. Malheureusement nous n'avons pas encore ce genre de produit à notre catalogue.

Je vous envoie séparément à cet email un devis pour une unité de ab21623.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

I see that:
Anti-acetyl Lysine antibody - ChIP Grade (ab21623)
Is available. Has it been used for Chromatin Immunoprecipitation of acetylated-transcription factor-DNA complex. Lets and example: I want to decipher the Acetylated vs non-acetylated transcription factors binding to the promoter of a gene. Example; binding of HFN4 (OR HNF3beta, CEBP etc)-acetylated to the promoter of a liver specific gene.

ANSWER:

Thank you for contacting Abcam.

The antibody ab21623 has been tested on acetylated histone, acetylated BSA, and acetylated MB and has no reaction to the non acetylated proteins. There are alsoafew references on our website that have used this antibody in ChIP and so you may be able to find more information there:

http://www.abcam.com/acetyl-Lysine-antibody-ChIP-Grade-ab21623-references.html



Please let me know if there is anything else I can help you with.

ab21623 acetyl Lysine antibody - ChIP Grade

Will this product recognize mono-, di-, and/or tri-acetylated proteins/

ANSWER:

Thank you for your patience in this matter.

The originator of ab21623 (acetyl Lysine antibody - ChIP Grade) has confirmed that this product recognizes mono, di, tri- and multi-acetylated proteins (on lysine residues). The more acetylated residues are in a protein (on lysine residues), the greater detection sensitivity.

I hope this information helps. Please don't hesitate to contact us again if you have further questions.