Revision 5

#25069Store at -20C

Cell Signaling Technology

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

WB, IP, ChIP, ChIP-seq, C&R, C&T

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q14781

Entrez-Gene Id:

84733

Product Information

Product Usage Information

For optimal ChIP results, use 5 μL of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:50
CUT&Tag 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

CBX2 (E3N6A) Rabbit mAb recognizes endogenous levels of total CBX2 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala100 of human CBX2 protein.

Background

The polycomb group (PcG) proteins contribute to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Chromobox protein homolog 2 (CBX2) is a core component of the PRC1 complex, which binds to the H3K27me3 mark and recruits PRC1 for transcriptional repression (7). CBX2 is frequently overexpressed in various cancers, including breast, ovarian, and AR-independent prostate cancer, where it is associated with poor prognosis (8-14). Furthermore, elevated CBX2 expression levels correlate with larger tumor size, increased lymph node metastasis, and positive human epidermal growth factor receptor 2 (HER-2) status in breast cancer (8). Mutations in CBX2 have also been attributed to 46,XY disorders of sex development (DSD), at least partially due to misregulation of the NR5A1/SF1 gene (15,16).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Park, I.K. et al. (2003) Nature 423, 302-5.
  4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
  5. Tavares, L. et al. (2012) Cell 148, 664-78.
  6. Gao, Z. et al. (2012) Mol Cell 45, 344-56.
  7. Kaustov, L. et al. (2011) J Biol Chem 286, 521-9.
  8. Chen, W.Y. et al. (2017) Oncol Lett 13, 1149-1156.
  9. Liang, Y.K. et al. (2017) Oncotarget 8, 92375-92387.
  10. Piqué, D.G. et al. (2019) Br J Cancer 120, 746-753.
  11. Zheng, S. et al. (2019) Am J Transl Res 11, 1668-1682.
  12. Iqbal, M.A. et al. (2021) Mol Oncol 15, 1450-1465.
  13. Wheeler, L.J. et al. (2018) Oncogenesis 7, 92.
  14. Clermont, P.L. et al. (2016) Clin Epigenetics 8, 16.
  15. Biason-Lauber, A. et al. (2009) Am J Hum Genet 84, 658-63.
  16. Sproll, P. et al. (2019) Sci Rep 9, 16552.

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 ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

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.
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.

Limited Uses

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