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25497
JARID1/KDM5 Histone Demethylase Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

JARID1/KDM5 Histone Demethylase Antibody Sampler Kit #25497

Citations (0)
Western blot analysis of extracts from MCF7, NCCIT, and COS-7 cell lines using Jarid1B (E2X6N) Rabbit mAb.
CUT&RUN was performed with K-562 cells and JARID1B (E2X6N) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding of JARID1B across GAPDH gene.
Western blot analysis of extracts from various cell lines using JARID1A (D28B10) Rabbit mAb.
Western blot analysis of extracts from NCCIT and F9 cell lines using JARID1C (D29B9) Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell lines using JARID1D (E4D4B) Rabbit mAb.
Immunoprecipitation of Jarid1B from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Jarid1B (E2X6N) Rabbit mAb. Western blot analysis was performed using Jarid1B (E2X6N) Rabbit mAb.
CUT&RUN was performed with K-562 cells and JARID1B (E2X6N) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding of JARID1B across chromosome 12 (upper), including GAPDH gene (lower).
Immunoprecipitation of JARID1D protein from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is JARID1D (E4D4B) Rabbit mAb. Western blot analysis was performed using JARID1D (E4D4B) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from K562 cells and Jarid1B (E2X6N) Rabbit 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 GAPDH, a known target gene of Jarid1B (see additional figure containing ChIP-qPCR data).
CUT&RUN was performed with K-562 cells and either JARID1B (E2X6N) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human γ-Actin Intron 3 Primers #5047 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.
Chromatin immunoprecipitations were performed with cross-linked chromatin from K562 cells and Jarid1B (E2X6N) Rabbit 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 chromosome 12 (upper), including GAPDH (lower), a known target gene of Jarid1B (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either Jarid1B (E2X6N) 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 GAPDH Exon 1 Primers #5516,SimpleChIP® Human γ-Actin Intron 3 Primers #5047, SimpleChIP® Human DHFR Intron 1 Primers #7531, 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.
To Purchase # 25497
Cat. # Size Qty. Price
25497T
1 Kit  (4 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
JARID1A (D28B10) Rabbit mAb 3876 20 µl
  • WB
  • IP
H M 200 Rabbit IgG
JARID1B (E2X6N) Rabbit mAb 15327 20 µl
  • WB
  • IP
  • ChIP
  • C&R
H M Mk 180 Rabbit IgG
JARID1C (D29B9) Rabbit mAb 5361 20 µl
  • WB
  • IP
H M 180 Rabbit IgG
JARID1D (E4D4B) Rabbit mAb 78995 20 µl
  • WB
  • IP
H Mk 190 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The JARID1/KDM5 Histone Demethylase Antibody Sampler Kit provides an economical means for detecting the JARID1/KDM5 family of proteins. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the JARID1/KDM5 Histone Demethylase Antibody Sampler Kit detects endogenous levels of its target protein. JARID1A (D28B10) Rabbit mAb detects endogenous levels of both isoforms of total JARID1A protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala274 of human JARID1A protein and Leu830 of human JARID1C protein, near the amino terminus of human JARID1B protein, and near the carboxy terminus of human JARID1D protein.

Background

The methylation state of lysine residues in histone proteins is a major determinant for formation of active and inactive regions of the genome and is crucial for proper programming of the genome during development (1,2). Jumonji C (JmjC) domain-containing proteins represent the largest class of potential histone demethylase proteins (3). 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 (3). Based on homology, both humans and mice contain at least 30 such proteins, which can be divided into 7 separate families (3). 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) (4). 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) (4). All four JARID proteins demethylate di- and tri-methyl histone H3 Lys4; JARID1B also demethylates mono-methyl histone H3 Lys4 (5-7). JARID1A is a critical RB-interacting protein and is required for Polycomb-Repressive Complex 2 (PRC2)-mediated transcriptional repression during ES cell differentiation (8). A JARID1A-NUP98 gene fusion is associated with myeloid leukemia (9). JARID1B, which interacts with many proteins including c-Myc and HDAC4, may play a role in cell fate decisions by blocking terminal differentiation (10-12). JARID1B is overexpressed in many breast cancers and may act by repressing multiple tumor suppressor genes, including BRCA1 and HOXA5 (13,14). 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 (7). JARID1C mutations are associated with X-linked mental retardation and epilepsy (15,16). JARID1D is uniquely localized to the Y chromosome, and functions as a tumor suppressor by repressing genes associated with cell invasiveness (17). JARID1D is frequently mutated in metastatic prostate tumors, and low JARID1D levels are associated with poor prognosis in prostate cancer patients (17).

  1. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop, 1-27.
  2. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
  3. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
  4. Benevolenskaya, E.V. (2007) Biochem Cell Biol 85, 435-43.
  5. Christensen, J. et al. (2007) Cell 128, 1063-76.
  6. Yamane, K. et al. (2007) Mol Cell 25, 801-12.
  7. Tahiliani, M. et al. (2007) Nature 447, 601-5.
  8. Pasini, D. et al. (2008) Genes Dev 22, 1345-55.
  9. van Zutven, L.J. et al. (2006) Genes Chromosomes Cancer 45, 437-46.
  10. Secombe, J. et al. (2007) Genes Dev 21, 537-51.
  11. Barrett, A. et al. (2007) Int J Cancer 121, 265-75.
  12. Dey, B.K. et al. (2008) Mol Cell Biol 28, 5312-27.
  13. Barrett, A. et al. (2002) Int J Cancer 101, 581-8.
  14. Lu, P.J. et al. (1999) J Biol Chem 274, 15633-45.
  15. Tzschach, A. et al. (2006) Hum Mutat 27, 389.
  16. Jensen, L.R. et al. (2005) Am J Hum Genet 76, 227-36.
  17. Li, N. et al. (2016) Cancer Res 76, 831-43.

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