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Ist ab24552 gegen NCoR1 oder 2? |
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ANSWER: |
Vielen Dank für Ihre Anfrage - und für Ihre Geduld! Bitte entschuldigen Sie, dass die Antwort so lange auf sich warten ließ. Ich hatte das Labor, das den NCoR- Antikörper ab24552 herstellt, kontaktiert und musste sehr lange auf eine zufriedenstellende Antwort warten. Ich kann Ihnen nun endlich mitteilen, dass ab24552 spezifisch für das humane NCoR1, Isoform 1 ist (Genbank Accession number AF044209, SwissProt ID O75376). Auf diese Sequenz bezieht sich die Angabe der Aminosäuren des Immunogens (789-957). Gerne kann ich Ihnen versichern, dass dieser Antikörper auch unserer Abcam Garantie unterliegt: Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und Sie sich innerhalb von sechs Monaten mit Ihrem Problem an uns wenden, werden wir Ihnen gerne Vorschläge zur Optimierung Ihres Protokolls oder gegebenenfalls einen Ersatz oder eine Gutschrift schicken. Ich hoffe, diese Informationen helfen Ihnen (auch jetzt noch) weiter und wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben. Normalerweise streben wir wesentlich schnellere Antworten an; es tut mir leid, dass das in diesem Fall nicht möglich war. |
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ORDER NUMBER 116253 the data sheet has no concentration on it so dont know how to use for the IP. i need to use 5micrograms
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ANSWER: |
I have just received information from the source of the antibody saying "We normally use 2-3 ul of the N-CoR antibody for IP. We normally use HeLa nuclear extracts for IP, although it also worked for whole cell extracts from HeLa or 293T. I attach a couple of papers in which this antibody was used for IP, western". The scientist refers to the references: Yoon HG et al. Reading and function of a histone code involved in targeting corepressor complexes for repression. Mol Cell Biol 25:324-35 (2005). PubMed: 15601853 Yoon HG et al. Purification and functional characterization of the human N-CoR complex: the roles of HDAC3, TBL1 and TBLR1. EMBO J 22:1336-46 (2003). PubMed: 12628926 I hope this information helps, please do not hesitate to contact me if I can be of further assistance, |
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What I wanted to know as far as positive control was something to the equivalent of "did they use rabbit whole serum or rabbit IgG as a control in the IP portion of the assay?" One has to use a positive control in this assay and if you look at those publications they show IgG in their figures. I am shocked that you don't test these internally in your ChIP system and rely solely on a customer's results - no internal standard testing is almost unbelievable.
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ANSWER: |
Thank you for your enquiry. We qualify our "ChIP grade" antibodies in three ways: Through our; 1. In our in-house ChIP testing program, 2. In a scientific publication or 3. by a customer review. On this occasion this antibody was sourced from the authors. We have every confidence in the behaviour of this antibody by ChIP. We make every effort to test as many of our antibodies in house as possible. However, this is not always possible given the cell systems employed to detect the binding of chromatin associated proteins. We only qualify our antibodies as being suitable for chromatin immunoprecipitation following a customers review if we are aware of the protocol used and the necessary optimisation for this antibody. We also monitor the status of our ChIP grade antibodies very closely. Should a ChIP grade antibody be shown to no longer demonstrate the capacity to immunoprecipitate cross linked chromatin we remove ChIP as an application. I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
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What was used as a positive control in testing these antibodies in ChIP? |
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ANSWER: |
Thank you for your enquiry. We qualify our antibodies as "ChIP grade" status in three ways 1. in our in-house ChIP testing program, 2. in a scientific publication and 3. by a customer (Abreview). Rabbit polyclonal to NCoR2, ab24551 was qualified as being capable of ChIP by its use in the following publication: Yoon HG et al. Reading and function of a histone code involved in targeting corepressor complexes for repression. Mol Cell Biol 25:324-35 (2005). PubMed: 15601853 In this publication they demonstrated the association of NCoR with the deiodinase 1 (D1) promoter (thyroid hormone response element (TRE)) in a HeLa a2 cell line which constitutively expresses a FLAG-tagged human TRa. They did not use a positive control as such. However, if this does not present itself as a suitable control any thyroid hormone controlled gene that is reactivated with Hdac inhibitors or has been shown to have association with NCoR are good places to start. ab24552 was also qualified by its presence in the same publication. It was used and confirmed by the same authors. I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
