Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Jarid2 antibody [EPR6357(2)] - ChIP Grade (ab192252)

Overview

  • Product name

    Anti-Jarid2 antibody [EPR6357(2)] - ChIP Grade
    See all Jarid2 primary antibodies
  • Description

    Rabbit monoclonal [EPR6357(2)] to Jarid2 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, ChIPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human Jarid2 aa 100-200. The exact sequence is proprietary.
    Database link: Q92833

  • Positive control

    • NCCIT, SH-SY5Y, 293 and U87-MG cell lysates; SH-SY5Y cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab192252 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 139 kDa (predicted molecular weight: 139 kDa).
ICC/IF 1/100.
ChIP Use at an assay dependent concentration.

Target

  • Function

    Regulator of histone methyltransferase complexes that plays an essential role in embryonic development, including heart and liver development, neural tube fusion process and hematopoiesis. Acts by modulating histone methyltransferase activity and promoting the recruitment of histone methyltransferase complexes to their target genes. Binds DNA and mediates the recruitment of the PRC2 complex to target genes in embryonic stem cells. Does not have histone demethylase activity but regulates activity of various histone methyltransferase complexes. In embryonic stem cells, it associates with the PRC2 complex and inhibits trimethylation of 'Lys-27' of histone H3 (H3K27me3) by the PRC2 complex, thereby playing a key role in differentiation of embryonic stem cells and normal development. In cardiac cells, it is required to repress expression of cyclin-D1 (CCND1) by activating methylation of 'Lys-9' of histone H3 (H3K9me) by the GLP1/EHMT1 and G9a/EHMT2 histone methyltransferases. Also acts as a transcriptional repressor of ANF via its interaction with GATA4 and NKX2-5. Participates in the negative regulation of cell proliferation signaling.
  • Tissue specificity

    During embryogenesis, predominantly expressed in neurons and particularly in dorsal root ganglion cells.
  • Sequence similarities

    Contains 1 ARID domain.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
  • Domain

    The ARID domain is required to target the PRC2 complex to its target genes.
    The GSGFP motif is required for the interaction with SUZ12.
  • Cellular localization

    Nucleus. Colocalizes with the PRC2 complex on chromatin.
  • Information by UniProt
  • Database links

  • Alternative names

    • JARD2 antibody
    • JARD2_HUMAN antibody
    • JARID2 antibody
    • JMJ antibody
    • Jumonji AT rich interactive domain 2 antibody
    • Jumonji homolog antibody
    • Jumonji like protein antibody
    • Jumonji protein antibody
    • Jumonji/ARID domain containing protein 2 antibody
    • Jumonji/ARID domain-containing protein 2 antibody
    • Protein Jumonji antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)              
    Lane 2: JARID2 (KO) knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: SH-SY5Y whole cell lysate (20 µg)
    Lane 4: U87MG whole cell lysate (20 µg)              

    Lanes 1 - 4: Merged signal (red and green). Green - ab192252 observed at 138 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab192252 was shown to specifically recognize JARID2 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when JARID2 knockout samples were examined. Wild-type and JARID2 knockout samples were subjected to SDS-PAGE.  Ab192252 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% triton X-100 permeabilized SH-SY5Y cells labeling Jarid2 with ab192252 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary antibody (ab150078) at 1/500 dilution. Nuclear counter stain Dapi (blue).

     

    The two negative controls are ab192252 at 1/100 dilution followed by Goat anti mouse IgG (Alexa Fluor®488) secondary antibody at 1/200 dilution.

  • Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab192252 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • All lanes : Anti-Jarid2 antibody [EPR6357(2)] - ChIP Grade (ab192252) at 1/10000 dilution

    Lane 1 : NCCIT cell lysate
    Lane 2 : SH-SY5Y cell lysate
    Lane 3 : 293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 139 kDa
    Observed band size: 139 kDa



    Blocking/Dilution buffer: 5% NFDM /TBST.

  • Anti-Jarid2 antibody [EPR6357(2)] - ChIP Grade (ab192252) at 1/1000 dilution + U87-MG cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 139 kDa
    Observed band size: 139 kDa



    Blocking/Dilution buffer: 5% NFDM /TBST.

References

This product has been referenced in:

  • Nguyen K  et al. Multiple Histone Lysine Methyltransferases Are Required for the Establishment and Maintenance of HIV-1 Latency. MBio 8:N/A (2017). WB, ChIP ; Human . Read more (PubMed: 28246360) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - whole cell (ESC)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
20 µg
Specification
ESC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C

Aurora Serrano

Verified customer

Submitted Mar 30 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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