Revision 3

#13768Store 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, ChIP, ChIP-seq

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

130

Source/Isotype:

Mouse IgG2b

UniProt ID:

#P78364

Entrez-Gene Id:

1911

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50
Chromatin IP-seq 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

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

Species Reactivity:

Human, Mouse

Source / Purification

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

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

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.

使用に関する制限

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Revision 3
#13768

PHC1 (1F3F3) Mouse mAb

Western Blotting Image 1: PHC1 (1F3F3) Mouse mAb Expand Image
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.
No image available
Chromatin Immunoprecipitation Image 1: PHC1 (1F3F3) Mouse mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated NCCIT cells and PHC1 (1F3F3) Mouse mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HoxA1, a known target gene of PHC1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: PHC1 (1F3F3) Mouse mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated NCCIT cells and PHC1 (1F3F3) Mouse mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across HoxA (upper) and HoxD (lower), known target genes of PHC1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: PHC1 (1F3F3) Mouse mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either PHC1 (1F3F3) Mouse mAb or 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.