Revision 10

#13594Store 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, C&R, C&T

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

150

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q92833

Entrez-Gene Id:

3720

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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 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

JARID2 (D6M9X) Rabbit mAb recognizes endogenous levels of total JARID2 protein.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp1114 of human JARID2 protein.

Background

Jumonji/ARID domain-containing protein 2 (JARID2) is a founding member of the JmjC-domain-containing protein family that is involved in regulation of histone methyltransferase activity (1,2). While many proteins in this family are protein demethylases, JARID2 lacks several conserved residues in the catalytic domain and does not exhibit protein demethylase activity (1,2). Research studies indicate that JARID2 is a nuclear protein that is highly expressed in poorly differentiated and actively dividing cells, with expression decreasing upon cellular differentiation (3,4). Expression of JARID2 protein is essential for embryonic development as the protein plays an important role in regulation of heart and liver development, neural tube fusion, and hematopoiesis (4). JARID2 is an accessory component of the polycomb repressor complex 2 (PRC2), which represses target gene expression through methylation of histone H3 at lysine 27 by EZH2 methyltransferase (5-10). JARID2 recruits the PRC2 complex to target genes and increases EZH2 methyltransferase activity by binding to nucleosomes and DNA (5-10). Additional studies show that loss of JARID2 expression results in decreased recruitment of PRC2, decreased methylation of histone H3 at lysine 27 at target genes, and delayed and incomplete differentiation of embryonic stem cells (5-10). Experimental knockdown of JARID2 in Xenopus laevis impairs the induction of gastrulation genes in blastula embryos and results in differentiation failure (5).

  1. Cloos, P.A. et al. (2008) Genes Dev 22, 1115-40.
  2. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
  3. Luzzani, C. et al. (2011) Biochem Biophys Res Commun 410, 816-22.
  4. Jung, J. et al. (2005) Dev Dyn 232, 21-32.
  5. Peng, J.C. et al. (2009) Cell 139, 1290-302.
  6. Shen, X. et al. (2009) Cell 139, 1303-14.
  7. Pasini, D. et al. (2010) Nature 464, 306-10.
  8. Li, G. et al. (2010) Genes Dev 24, 368-80.
  9. Son, J. et al. (2013) Genes Dev 27, 2663-77.
  10. Kaneko, S. et al. (2014) Mol Cell 53, 290-300.

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 nonfat dry milk, 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 C&R: CUT&RUN C&T: CUT&Tag

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.

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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 10
#13594

JARID2 (D6M9X) Rabbit mAb

Western Blotting Image 1: JARID2 (D6M9X) Rabbit mAb Expand Image
Western blot analysis of extracts from F9 cells and NTERA-2 cells, untreated (-) or treated with retinoic acid (1 μM, 5 d; +) to induce differentiation, using JARID2 (D6M9X) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, JARID2 protein levels decrease upon NTERA-2 cell differentiation.
No image available
Chromatin Immunoprecipitation Image 1: JARID2 (D6M9X) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either JARID2 (D6M9X) Rabbit mAb, CBX8 (D2O8C) Rabbit mAb #14696, or AEBP2 (D7C6X) Rabbit mAb #14129, 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. All of JARID2, CBX8, and AEBP2 are known to associate with PRC complexes. The figure shows binding of JARID2, CBX8, and AEBP2 across HOXD gene region, known target genes of JARID2, CBX8, and AEBP2 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: JARID2 (D6M9X) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either JARID2 (D6M9X) Rabbit mAb, CBX8 (D2O8C) Rabbit mAb #14696, or AEBP2 (D7C6X) Rabbit mAb #14129, 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. All of JARID2, CBX8, and AEBP2 are known to associate with PRC complexes. The figure shows binding of JARID2, CBX8, and AEBP2 across HOXA (upper) and HOXD (lower), known target genes of JARID2, CBX8, and AEBP2 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: JARID2 (D6M9X) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either JARID2 (D6M9X) 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 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.
CUT and RUN Image 1: JARID2 (D6M9X) Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and JARID2 (D6M9X) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HoxA, a known target gene of JARID2 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: JARID2 (D6M9X) Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and JARID2 (D6M9X) Rabbit mAb, using CUT&RUN Assay Kit #86652. 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 JARID2 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: JARID2 (D6M9X) Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and either JARID2 (D6M9X) 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 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.
CUT and Tag Image 1: JARID2 (D6M9X) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and JARID2 (D6M9X) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across HOXA1, a known target gene of JARID2 (see our ChIP-qPCR figure).
CUT and Tag Image 2: JARID2 (D6M9X) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and JARID2 (D6M9X) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across the HOXA gene cluster (upper), and the HOXD gene cluster (lower), which are known target genes of JARID2 (see our ChIP-qPCR figure).
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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.