Overview

  • Product name
    Anti-Nuclear Receptor Corepressor NCoR antibody
    See all Nuclear Receptor Corepressor NCoR primary antibodies
  • Description
    Rabbit polyclonal to Nuclear Receptor Corepressor NCoR
  • Host species
    Rabbit
  • Specificity
    This antibody is specific to NCoR1,variant 1, Genbank Accession number AF044209, SwissProt ID O75376.
  • Tested applications
    Suitable for: ChIP, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Monkey
  • Immunogen

    Recombinant fragment, corresponding to amino acids 789-957 of Human Nuclear Receptor Corepressor NCoR

  • Positive control
    • HeLa nuclear lysate

Applications

Our Abpromise guarantee covers the use of ab24552 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent dilution.
IP Use at an assay dependent dilution. Use 2-3ul per IP (See references for protocol details).
WB 1/1000. Detects a band of approximately 270 kDa (predicted molecular weight: 270 kDa).
ICC/IF Use at an assay dependent dilution.

Target

  • Function
    Mediates transcriptional repression by certain nuclear receptors. Part of a complex which promotes histone deacetylation and the formation of repressive chromatin structures which may impede the access of basal transcription factors.
  • Sequence similarities
    Belongs to the N-CoR nuclear receptor corepressors family.
    Contains 2 SANT domains.
  • Domain
    The N-terminal region contains three independent domains that are capable of mediating transcriptional repression (RD1, RD2 and RD3).
    The C-terminal region contains two separate nuclear receptor-interacting domains (ID1 and ID2), each of which contains a conserved sequence referred to as the CORNR box. This motif is necessary and sufficient for binding to unligated nuclear hormone receptors, while sequences flanking the CORNR box determine the precise nuclear hormone receptor specificity.
  • Post-translational
    modifications
    Ubiquitinated; mediated by SIAH2 and leading to its subsequent proteasomal degradation.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • hN CoR antibody
    • hNCoR antibody
    • KIAA1047 antibody
    • N CoR antibody
    • N Cor/SMRT corepressor Rip13 antibody
    • N CoR1 antibody
    • N-CoR antibody
    • N-CoR1 antibody
    • NCOR 1 antibody
    • NCoR antibody
    • Ncor1 antibody
    • NCOR1_HUMAN antibody
    • Nuclear receptor co repressor 1 antibody
    • Nuclear receptor corepressor 1 antibody
    • Retinoid X receptor interacting protein 13 antibody
    • RIP13 antibody
    • Rxrip13 antibody
    • thyroid hormone and retinoic acid receptor associated corepressor 1 antibody
    • TRAC 1 antibody
    • TRAC1 antibody
    see all

Images

  • Anti-Nuclear Receptor Corepressor NCoR antibody (ab24552) at 1/1000 dilution + HeLa nuclear extract

    Predicted band size: 270 kDa
    Observed band size: 270 kDa

References

This product has been referenced in:
  • Chakraborty K  et al. The mito-DAMP cardiolipin blocks IL-10 production causing persistent inflammation during bacterial pneumonia. Nat Commun 8:13944 (2017). Read more (PubMed: 28074841) »
  • Viswakarma N  et al. Transcriptional regulation of mixed lineage kinase 3 by estrogen and its implication in ER-positive breast cancer pathogenesis. Oncotarget 8:33172-33184 (2017). ChIP . Read more (PubMed: 28388540) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Rat Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 07 2014

Application
ChIP
Sample
Human Cell lysate - nuclear (MCF7)
Specification
MCF7
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: FORMALDEHYDE 10%

Abcam user community

Verified customer

Submitted Dec 10 2013

Answer

Thank you for your reply.

I am sorry the results you have observed have not improved with ab24552. From looking at the results you have shared you appear to be getting very similar results, with ab24552 showing a much clearer band at ˜250 kDa than with ab58396. As discussed previously, the band at ˜250 kDa may well be the band that you are expecting. Although the SwissProt reference for the Nuclear Receptor Corepressor NCoR states that isoforms of ˜260 and 270 have been observed of this protein. The antibodies you have used have both been raised against regions of the NCoR protein which are present in both isoforms so if either or both of the proteins are expressed in your samples they would be expected to be detected. I have not been able to find any further information as to which of the isoforms is most prevalent. I would therefore not be able offer a suggestion as to which of the two isoforms it may be that you are detecting.

As the results you are seeing with both antibodies are consistent (both show bands of ˜250 kDa), I would suggests this supports this being a specific band. If you would like to confirm this further you could isolate the protein in the band and perform Mass Spec analysis. Alternatively, if you have access to a construct which can be used to over-express the protein in a cell line this can often prove a useful positive and negative control.

You are seeing a shift of about 20 kDa in the size of the protein on your membrane. This size of shift is not uncommon for proteins of this size. Separation by SDS-PAGE does not give the exact size of proteins. It relies on the proteins being coated in SDS proportional to their size, providing a negative charge proportional to their size. This is not always the case. The degree of SDS binding the protein can be affected by its hydrophobicity, charge, glycosylation, phosphorylation, or simply the amino acid sequence. This can also be different depending on the lysis buffer and running buffers used when running the gel.

I hope this information has been of help. If you would like for me to help optimise the results, if you could possibly provide me with more details of how you are running the experiment now. I have attached our questionnaire for this reason. Could you also detail how the tissue is being processed (which lysis buffer and the running buffers used)?

I look forward to receiving your reply.

Read More

Question
Answer

Thank you for contacting Abcam earlier today.
As we discussed, I am sending you new aliquots of both ab24552 and ab26721 and your new order number is ***********.
Below are a few possible alternatives if the replacements are not successful:
- https://www.abcam.com/Nuclear-Receptor-Corepressor-NCoR-antibody-ChIP-Grade-ab3482.html
- https://www.abcam.com/RNA-polymerase-II-CTD-repeat-YSPTSPS-antibody-8WG16-ChIP-Grade-ab817.html
- https://www.abcam.com/RNA-polymerase-II-CTD-repeat-YSPTSPS-antibody-H14-ChIP-Grade-ab24759.html
Please let me know if there is anything else I can help you with.

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Question
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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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (vero)
Gel Running Conditions
Reduced Denaturing
Loading amount
20000 cells
Specification
vero
Blocking step
Milk as blocking agent for 1 minute(s) · Concentration: 0.1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 23 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (MCF-7 Breast Cancer cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
MCF-7 Breast Cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Dr. Corina Marx

Verified customer

Submitted Jul 20 2007

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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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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Question
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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