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  1. Link

    chd3-antibody-epncir110a-ab109195.pdf

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Immunology Immune System Diseases Autoimmune
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CHD3 antibody [EPNCIR110A] (ab109195)

  • Datasheet
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Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Flow Cytometry (Intracellular) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Immunocytochemistry/ Immunofluorescence - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
  • Anti-CHD3 antibody [EPNCIR110A] (ab109195)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPNCIR110A] to CHD3
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-CHD3 antibody [EPNCIR110A]
    See all CHD3 primary antibodies
  • Description

    Rabbit monoclonal [EPNCIR110A] to CHD3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

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

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • K562, HeLa, C6, and PC12 cell lysates, Human breast and Human ovary tissues; HeLa cells
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Gordon Hager. View antibodies from NCI Center for Cancer Research Collaboration.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPNCIR110A
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Immune System Diseases
    • Autoimmune
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Mi-2
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors

Associated products

  • Alternative Versions

    • Anti-CHD3 antibody [EPNCIR110A] - BSA and Azide free (ab240942)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • PC-12 nuclear extract lysate (ab14884)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109195 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 260 kDa (predicted molecular weight: 227 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)
ICC/IF
1/100 - 1/250.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 260 kDa (predicted molecular weight: 227 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for IP.

Target

  • Function

    Component of the histone deacetylase NuRD complex which participates in the remodeling of chromatin by deacetylating histones. Required for anchoring centrosomal pericentrin in both interphase and mitosis, for spindle organization and centrosome integrity.
  • Tissue specificity

    Widely expressed.
  • Sequence similarities

    Belongs to the SNF2/RAD54 helicase family.
    Contains 2 chromo domains.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 2 PHD-type zinc fingers.
  • Cellular localization

    Nucleus. Cytoplasm > cytoskeleton > centrosome. Associates with centrosomes in interphase and mitosis.
  • Target information above from: UniProt accession Q12873 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1107 Human
    • Entrez Gene: 216848 Mouse
    • Entrez Gene: 303241 Rat
    • Omim: 602120 Human
    • SwissProt: Q12873 Human
    • Unigene: 25601 Human
    • Alternative names

      • 226 antibody
      • 2600010P09Rik antibody
      • AF020312 antibody
      • ATP dependent helicase CHD3 antibody
      • ATP-dependent helicase Chd3 antibody
      • CHD-3 antibody
      • Chd3 antibody
      • CHD3_HUMAN antibody
      • Chd7 antibody
      • Chromodomain-helicase-DNA-binding protein 3 antibody
      • hZFH antibody
      • MGC40857 antibody
      • Mi 2 autoantigen 240 kDa protein antibody
      • Mi 2a antibody
      • Mi-2 autoantigen 240 kDa protein antibody
      • Mi2 ALPHA antibody
      • Mi2-alpha antibody
      • Prp7 antibody
      • Prp9 1 antibody
      • ZFH antibody
      • Zinc finger helicase antibody
      • zinc-finger helicase (Snf2-like) antibody
      see all

    Images

    • Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: CHD3 knockout HAP1 cell lysate (20 µg)
      Lane 3: Raji cell lysate (20 µg)
      Lane 4: Y79 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109195 observed at 245 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109195 was shown to specifically react with CHD3 when CHD3 knockout samples were used. Wild-type and CHD3 knockout samples were subjected to SDS-PAGE. ab109195 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW)preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)

      Immunohistochemical staining of paraffin-embedded Human breast tissue using ab109195 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Flow Cytometry (Intracellular) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling CHD3 with purified ab109195 at 1/230 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Western blot - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      All lanes : Anti-CHD3 antibody [EPNCIR110A] (ab109195) at 1/1000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : C6 cell lysate
      Lane 4 : PC12 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 227 kDa
      Observed band size: 260 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD3 antibody [EPNCIR110A] (ab109195)

      Immunohistochemical staining of paraffin-embedded Human ovary tissue using ab109195 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Immunocytochemistry/ Immunofluorescence - Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Immunofluorescent staining of HeLa cells using ab109195 at a dilution of 1/100.
    • Anti-CHD3 antibody [EPNCIR110A] (ab109195)
      Anti-CHD3 antibody [EPNCIR110A] (ab109195)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (2)

    Publishing research using ab109195? Please let us know so that we can cite the reference in this datasheet.

    ab109195 has been referenced in 2 publications.

    • Xie J  et al. The chromatin-remodeling enzyme CHD3 plays a role in embryonic viability but is dispensable for early vascular development. PLoS One 15:e0235799 (2020). PubMed: 32658897
    • Rodríguez-Castañeda F  et al. The SUMO protease SENP1 and the chromatin remodeler CHD3 interact and jointly affect chromatin accessibility and gene expression. J Biol Chem 293:15439-15454 (2018). PubMed: 30082317

    Customer reviews and Q&As

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