Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

PHC1 (1F3F3) Mouse mAb #13768

No. Size Price
13768S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
13768 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 130 Mouse IgG2b
IP 1:50
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Specificity / Sensitivity

PHC1 (1F3F3) Mouse mAb recognizes endogenous levels of total PHC1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human PHC1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PHC1 (1F3F3) Mouse mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, PHC1 protein is not expressed in DU 145 or NIH/3T3 cells.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 µl of PHC1 (1F3F3) Mouse mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, 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.

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). Polycomb group proteins regulate cell proliferation and senescence through repression of the p16Ink4a and p19Arf genes, and are essential in maintaining adult hematopoietic, neural stem cells, and embryonic stem cells (3-5). PcG proteins are found in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. DNA-binding transcription factors recruit the EED-EZH2 complex to genes, which methylates histone H3 on Lys27 (6). Methylation of Lys27 facilitates the recruitment of the PRC1 complex, which ubiquitinylates histone H2A on Lys119 (7). PRC1 is composed of BMI1 and RING1A, which enhance the E3 ubiquitin ligase activity of the RING1B catalytic subunit (8). Polyhomeotic-like 1 (PHC1) is one of several additional PRC1 complex proteins that are required to maintain the silenced state of PRC1 target genes and mediate proper anterior-posterior specification during development (9). Mutations in the corresponding PHC1 gene correlate with an autosomal recessive form of primary microcephaly characterized by low-to-normal cognitive function and impaired DNA repair (10).

  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. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
  6. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  7. Wang, H. et al. (2004) Nature 431, 873-8.
  8. Cao, R. et al. (2005) Mol Cell 20, 845-54.
  9. Isono, K. et al. (2005) Mol Cell Biol 25, 6694-706.
  10. Awad, S. et al. (2013) Hum Mol Genet 22, 2200-13.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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