Revision 1

#34271Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

270

Source/Isotype:

Rabbit IgG

UniProt ID:

#O75376

Entrez-Gene Id:

9611

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 10^6 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:100
Immunofluorescence (Immunocytochemistry) 1:800
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

NCoR1 (E4N4S) Rabbit mAb recognizes endogenous levels of total NCoR1 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human NCoR1 protein.

Background

The most well characterized nuclear receptor corepressors are SMRT (silencing mediator for retinoic acid and thyroid hormone receptors) and its close paralog NCoR1 (nuclear receptor corepressor) (1,2). NCoR1 functions to transcriptionally silence various unliganded, DNA bound non-steroidal nuclear receptors by serving as a large molecular scaffold that bridges the receptors with multiple chromatin remodeling factors that repress nuclear receptor-mediated gene transcription, in part, through deacetylation of core histones surrounding target promoters. Indeed, the N-terminal portion of NCoR1 possesses multiple distinct transcriptional repression domains (RDs) reponsible for the recruitment of additional components of the corepressor complex such as HDACs, mSin3, GPS2, and TBL1/TBLR1. In between the RDs lies a pair of potent repressor motifs known as SANT motifs (SWI3, ADA2, N-CoR, and TFIIIB), which recruit HDAC3 and histones to the repressor complex in order to enhance HDAC3 activity (3). The C-terminal portion of NCoR1 contains multiple nuclear receptor interaction domains (NDs), each of which contains a conserved CoRNR box (or L/I-X-X-I/V-I) motif that allow for binding to various unliganded nuclear hormone receptors such as thyroid hormone (THR) and retinoic acid (RAR) receptors (4,5).

Recent genetic studies in mice have not only corroborated the wealth of biochemical studies involving NCoR1 but have also provided significant insight regarding the function of NCoR1 in mammalian development and physiology. Although it has been observed that loss of Ncor1 does not affect early embyonic development, likely due to compensation by Smrt, embryonic lethality ultimately results during mid-gestation, largely due to defects in erythropoesis and thymopoesis (6). Another study demonstrated that the NDs of NCoR1 are critical for its ability to function in a physiological setting as a transcriptional repressor of hepatic THR and Liver X Receptor (LXR) (7).

  1. Chen, J.D. and Evans, R.M. (1995) Nature 377, 454-7.
  2. Hörlein, A.J. et al. (1995) Nature 377, 397-404.
  3. Jones, P.L. and Shi, Y.B. (2003) Curr Top Microbiol Immunol 274, 237-68.
  4. Downes, M. et al. (1996) Nucleic Acids Res 24, 4379-86.
  5. Wong, C.W. and Privalsky, M.L. (1998) Mol Cell Biol 18, 5724-33.
  6. Jepsen, K. et al. (2000) Cell 102, 753-63.
  7. Astapova, I. et al. (2008) Proc Natl Acad Sci U S A 105, 19544-9.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 1
#34271

NCoR1 (E4S4N) Rabbit mAb

Western Blotting Image 1: NCoR1 (E4S4N) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using NCoR1 (E4N4S) Rabbit mAb. As expected, HCT 116 cells are low for NCoR1 expression.
Immunoprecipitation Image 1: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunoprecipitation of NCoR1 from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NCoR1 (E4N4S) Rabbit mAb. Western blot analysis was performed using NCoR1 (E4N4S) Rabbit mAb.
Immunohistochemistry Image 1: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using NCoR1 (E4S4N) Rabbit mAb.
Immunohistochemistry Image 2: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded ACHN cell pellet (left, high-expressing) or HCT 116 cell pellet (right, low-expressing) using NCoR1 (E4S4N) Rabbit mAb.
Immunohistochemistry Image 3: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using NCoR1 (E4S4N) Rabbit mAb.
Immunohistochemistry Image 4: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using NCoR1 (E4S4N) Rabbit mAb.
Immunohistochemistry Image 5: NCoR1 (E4S4N) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using NCoR1 (E4S4N) Rabbit mAb.
Immunofluorescence Image 1: NCoR1 (E4S4N) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of ACHN cells (left, high-expressing) or HCT 116 cells (right, low-expressing) using NCOR1 (E4S4N) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Nuclei were labeled with DAPI #4083 (blue).
Chromatin Immunoprecipitation Image 1: NCoR1 (E4S4N) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from LS 180 cells treated with vitamin D (10 nM) for 3 hours and either NCoR1 (E4S4N) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Upstream Primers #25661, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.