Revision 5

#15327

Store at -20C

JARID1B (E2X6N) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, ChIP, ChIP-seq, C&R

Reactivity:
H M Mk

Sensitivity:
Endogenous

MW (kDa):
180

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9UGL1

Entrez-Gene Id:
10765

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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 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

Jarid1B (E2X6N) Rabbit mAb recognizes endogenous levels of total Jarid1B protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Jarid1B 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).

Background References

  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.

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

W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: Human M: Mouse Mk: Monkey

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

#15327

JARID1B (E2X6N) Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from MCF7, NCCIT, and COS-7 cell lines using Jarid1B (E2X6N) Rabbit mAb.
Western Blotting Image 1: JARID1B (E2X6N) 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.
Immunoprecipitation Image 1: JARID1B (E2X6N) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

#15327

JARID1B (E2X6N) Rabbit mAb

Cell Signaling Technology Logo
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).
Chromatin Immunoprecipitation Image 1: JARID1B (E2X6N) Rabbit mAb
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 Immunoprecipitation Image 2: JARID1B (E2X6N) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

#15327

JARID1B (E2X6N) Rabbit mAb

Cell Signaling Technology Logo
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.
Chromatin Immunoprecipitation Image 3: 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.
CUT & RUN Image 1: 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).
CUT & RUN Image 2: JARID1B (E2X6N) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

#15327

JARID1B (E2X6N) Rabbit mAb

Cell Signaling Technology Logo
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.
CUT & RUN Image 3: JARID1B (E2X6N) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.