Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Jumonji Family Antibody Sampler Kit #8340

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
JARID1A (D28B10) XP® Rabbit mAb #3876 40 µl W IP IF-IC H M (R) (B) 200 Rabbit IgG
JARID1B Antibody #3273 40 µl W IP H Mk 180 Rabbit
JARID1C (D29B9) Rabbit mAb #5361 40 µl W IP H M (Mk) (Pg) (Hr) 180 Rabbit IgG
JMJD2A (C37E5) Rabbit mAb #5328 40 µl W IP IHC-P IF-IC H M R Mk 150 Rabbit IgG
JMJD2B Antibody #2898 40 µl W IP H (Mk) 150 Rabbit
JMJD1B (6A1-1F5) Mouse mAb #5377 40 µl W IP IF-IC H M R Mk 220 Mouse IgG1
PHF2 (D45A2) Rabbit mAb #3497 40 µl W IP H M R Mk 150 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl Horse

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Pg=Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Jumonji Family Antibody Sampler Kit detects endogenous levels of the respective target protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using JMJD2B Antibody #2898.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using JARID1B Antibody #3273.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PHF2 (D45A2) Rabbit mAb #3497.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using JARID1A (D28B10) XP® Rabbit mAb #3876.

Western Blotting

Western Blotting

Western blot analysis of extracts from NCCIT and NTERA-2 cells using JMJD2A (C37E5) Rabbit mAb #5328.

Western Blotting

Western Blotting

Western blot analysis of extracts from NCCIT and F9 cells using JARID1C (D29B9) Rabbit mAb #5361.


Western Blotting

Western Blotting

Western blot analysis of extracts from NCCIT cells using JMJD1B (6A1-1F5) Mouse mAb #5377.

Description

The Jumonji Family Antibody Sampler Kit provides an economical means of evaluating total levels of Jumonji family proteins. The kit contains enough primary antibodies to perform four western blot experiments with each primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to human JARID1A protein, residues surrounding Leu830 of the human JARID1C protein, residues surrounding Val444 of human JMJD2A protein, or human PHF2 protein. Monoclonal antibodies are produced by immunizing animals with a recombinant protein derived from the sequence of human JMJD1B protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to human JARID1B protein or human JMJD2B protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Jumonji C (JmjC) domain-containing proteins represent the largest class of potential histone demethylase proteins (1). The JmjC domain can catalyze the demethylation of mono-, di-, and tri-methyl lysine residues via an oxidative reaction that requires iron and α-ketoglutarate (1). Based on homology, both humans and mice contain at least 30 such proteins, which can be divided into 7 separate families (1).The JMJD2 (Jumonji domain-containing protein 2) family, also known as JHDM3 (JmjC domain-containing histone demethylation protein 3) family, contains four members: JMJD2A/JHDM3A, JMJD2B/JHDM3B, JMJD2C/JHDM3C, and JMJD2D/JHDM3D. In addition to the JmjC domain, these proteins also contain JmjN, PHD and Tudor domains, the latter of which has been shown to bind to methylated histone H3 at Lys4 and Lys9, and methylated histone H4 at Lys20 (2,3). JMJD2 proteins have been shown to demethylate di- and tri-methyl histone H3 at Lys9 and Lys36, and function as both activators and repressors of transcription (4-9). JMJD2A, JMJD2C and JMJD2D function as coactivators of the androgen receptor in prostate tumor cells (5). In contrast, JMJD2A also associates with Rb and N-CoR corepressor complexes and is necessary for transcriptional repression of target genes (6,7). JMJD2B antagonizes histone H3 Lys9 tri-methylation at pericentric heterochromatin (8). JMJD1B is a more widely expressed family member and is frequently deleted in myeloid leukemia (10).The JARID (Jumonji/AT-rich interactive domain-containing protein) family contains four members: JARID1A (also RBP2 and RBBP2), JARID1B (also PLU-1), JARID1C (also SMCX), and JARID1D (also SMCY) (11). In addition to the JmjC domain, these proteins contain JmjN, BRIGHT, C5HC2 zinc-finger, and PHD domains, the latter of which binds to methylated histone H3 (Lys9) (11). All four JARID proteins demethylate di- and tri-methyl histone H3 Lys4; JARID1B also demethylates mono-methyl histone H3 Lys4 (12-14). JARID1A is a critical RB-interacting protein and is required for Polycomb-Repressive Complex 2 (PRC2)-mediated transcriptional repression during ES cell differentiation (15). A JARID1A-NUP98 gene fusion is associated with myeloid leukemia (16). JARID1B, which interacts with many proteins including c-Myc and HDAC4, may play a role in cell fate decisions by blocking terminal differentiation (17-19). JARID1B is over-expressed in many breast cancers and may act by repressing multiple tumor suppressor genes including BRCA1 and HOXA5 (20,21). JARID1C has been found in a complex with HDAC1, HDAC2, G9a, and REST, which binds to and represses REST target genes in non-neuronal cells (14). JARID1D is largely uncharacterized. PHF2 contains a JmjC domain, which may play a role in histone demethylation (1).

  1. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
  2. Chen, Z. et al. (2007) Proc Natl Acad Sci U S A 104, 10818-23.
  3. Lee, J. et al. (2008) Nat Struct Mol Biol 15, 109-11.
  4. Whetstine, J.R. et al. (2006) Cell 125, 467-81.
  5. Shin, S. and Janknecht, R. (2007) Biochem Biophys Res Commun 359, 742-6.
  6. Gray, S.G. et al. (2005) J Biol Chem 280, 28507-18.
  7. Zhang, D. et al. (2005) Mol Cell Biol 25, 6404-14.
  8. Fodor, B.D. et al. (2006) Genes Dev 20, 1557-62.
  9. Cloos, P.A. et al. (2006) Nature 442, 307-11.
  10. Hu, Z. et al. (2001) Oncogene 20, 6946-54.
  11. Benevolenskaya, E.V. (2007) Biochem Cell Biol 85, 435-43.
  12. Christensen, J. et al. (2007) Cell 128, 1063-76.
  13. Yamane, K. et al. (2007) Mol Cell 25, 801-12.
  14. Tahiliani, M. et al. (2007) Nature 447, 601-5.
  15. Pasini, D. et al. (2008) Genes Dev 22, 1345-55.
  16. van Zutven, L.J. et al. (2006) Genes Chromosomes Cancer 45, 437-46.
  17. Secombe, J. et al. (2007) Genes Dev 21, 537-51.
  18. Barrett, A. et al. (2007) Int J Cancer 121, 265-75.
  19. Dey, B.K. et al. (2008) Mol Cell Biol 28, 5312-27.
  20. Barrett, A. et al. (2002) Int J Cancer 101, 581-8.
  21. Lu, P.J. et al. (1999) J Biol Chem 274, 15633-45.

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