Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ARID1A antibody [EPR13501] (ab182560)

Submit a review Q&A (1)References (13)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Western blot - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Flow Cytometry - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)
  • Anti-ARID1A antibody [EPR13501] (ab182560)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13501] to ARID1A
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ARID1A antibody [EPR13501]
    See all ARID1A primary antibodies

  • Description

    Rabbit monoclonal [EPR13501] to ARID1A

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details

  • Species reactivity

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

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human kidney and human adenocarcinoma of endometrium without ARID1A mutation tissues. ICC/IF: Wildtype HAP1 and SH-SY5Y cells. WB: HEK-293T and SH-SY5Y cell lysates.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab182560 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
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
WB
1/1000. Predicted molecular weight: 242 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.
Notes
Flow Cyt
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
WB
1/1000. Predicted molecular weight: 242 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.

Target

  • Function

    Involved in transcriptional activation and repression of select genes by chromatin remodeling (alteration of DNA-nucleosome topology). Binds DNA non-specifically. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene. Belongs to the neural progenitors-specific chromatin remodeling complex (npBAF complex) and the neuron-specific chromatin remodeling complex (nBAF complex). During neural development a switch from a stem/progenitor to a post-mitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state. The transition from proliferating neural stem/progenitor cells to post-mitotic neurons requires a switch in subunit composition of the npBAF and nBAF complexes. As neural progenitors exit mitosis and differentiate into neurons, npBAF complexes which contain ACTL6A/BAF53A and PHF10/BAF45A, are exchanged for homologous alternative ACTL6B/BAF53B and DPF1/BAF45B or DPF3/BAF45C subunits in neuron-specific complexes (nBAF). The npBAF complex is essential for the self-renewal/proliferative capacity of the multipotent neural stem cells. The nBAF complex along with CREST plays a role regulating the activity of genes essential for dendrite growth.

  • Tissue specificity

    Highly expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon, and PBL, and at a much lower level in heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas.

  • Sequence similarities

    Contains 1 ARID domain.

  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • actin-dependent regulator of chromatin subfamily F member 1 antibody
      • ARI1A_HUMAN antibody
      • ARID domain containing protein 1A antibody
      • ARID domain-containing protein 1A antibody
      • ARID1A antibody
      • AT rich interactive domain 1A (SWI like) antibody
      • AT rich interactive domain 1A antibody
      • AT rich interactive domain containing protein 1A antibody
      • AT-rich interactive domain-containing protein 1A antibody
      • B120 antibody
      • BAF250 antibody
      • BAF250A antibody
      • BM029 antibody
      • brain protein 120 antibody
      • BRG1 associated factor 250 antibody
      • BRG1 associated factor 250a antibody
      • BRG1-associated factor 250 antibody
      • BRG1-associated factor 250a antibody
      • C1ORF4 antibody
      • chromatin remodeling factor p250 antibody
      • chromosome 1 open reading frame 4 antibody
      • ELD antibody
      • hELD antibody
      • hOSA1 antibody
      • matrix-associated antibody
      • MRD14 antibody
      • Osa homolog 1 antibody
      • OSA1 antibody
      • OSA1 nuclear protein antibody
      • P270 antibody
      • SMARCF1 antibody
      • SWI like protein antibody
      • SWI SNF complex protein p270 antibody
      • SWI-like protein antibody
      • SWI/SNF complex protein p270 antibody
      • SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily f, member 1 antibody
      • SWI/SNF-related antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)

      Immunohistochemical analysis of paraffin embedded Human kidney tissue (Left image) labeling ARID1A using ab182560 at 1/1000 dilution. Right image: Right picture: paraffine embedded human clear cell carcinoma of kidney with ARID1A mutation. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA, pH 9 was performed before commencing with IHC staining protocol.

    • Western blot - Anti-ARID1A antibody [EPR13501] (ab182560)
      Western blot - Anti-ARID1A antibody [EPR13501] (ab182560)
      All lanes : Anti-ARID1A antibody [EPR13501] (ab182560) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : ARID1A knockout HEK-293T cell lysate
      Lane 3 : SH-SY5Y cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 242 kDa
      Observed band size: 270 kDa why is the actual band size different from the predicted?



      Lanes 1 - 3: Merged signal (red and green). Green - ab182560 observed at 270 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab182560 was shown to react with ARID1A in wild-type HEK-293T cells in Western blot with loss of signal observed in ARID1A knockout cell line ab266189 (ARID1A knockout cell lysate ab257250). Wild-type HEK-293T and ARID1A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab182560 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)
      Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)

      ab182560 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182560 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Flow Cytometry - Anti-ARID1A antibody [EPR13501] (ab182560)
      Flow Cytometry - Anti-ARID1A antibody [EPR13501] (ab182560)

      Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling ARID1A  with purified ab182560 at 1/230 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)
      Immunocytochemistry/ Immunofluorescence - Anti-ARID1A antibody [EPR13501] (ab182560)

      Immunofluorescent analysis of SH-SY5Y cells labeling ARID1A with ab182560 at 1/500 and Goat anti rabbit IgG(Alexa Fluor®555) at 1/200. Image at the right stained with DAPI.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] (ab182560)

      Immunohistochemical analysis of paraffin embedded Human adenocarcinoma of endometrium without ARID1A mutation (Left image) labeling ARID1A using ab182560 at 1/1000 dilution. Right image: Right picture: paraffine embedded human adenocarcinoma of endometrium with ARID1A mutation. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA, pH 9 was performed before commencing with IHC staining protocol.

    • Anti-ARID1A antibody [EPR13501] (ab182560)
      Anti-ARID1A antibody [EPR13501] (ab182560)

    References (13)

    Publishing research using ab182560? Please let us know so that we can cite the reference in this datasheet.

    ab182560 has been referenced in 13 publications.

    • Hackeng WM  et al. Alternative Lengthening of Telomeres and Differential Expression of Endocrine Transcription Factors Distinguish Metastatic and Non-metastatic Insulinomas. Endocr Pathol 31:108-118 (2020). PubMed: 32103422
    • Raab JR  et al. SWI/SNF remains localized to chromatin in the presence of SCHLAP1. Nat Genet 51:26-29 (2019). PubMed: 30510238
    • Han L  et al. Chromatin remodeling mediated by ARID1A is indispensable for normal hematopoiesis in mice. Leukemia 33:2291-2305 (2019). PubMed: 30858552
    • Wu S  et al. ARID1A spatially partitions interphase chromosomes. Sci Adv 5:eaaw5294 (2019). PubMed: 31131328
    • Zhao B  et al. ARID1A promotes genomic stability through protecting telomere cohesion. Nat Commun 10:4067 (2019). PubMed: 31492885
    View all Publications for this product

    Customer reviews and Q&As

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    Question

    Inquiry: What is the difference between ab182561 and ab182560

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    Answer

    These two antibodies are extremely similar. They recognise the same protein in the same applications and species however they are different clones. The immunogen used for both antibodies is the same (aa1200-1350).

    Although we have not mapped the epitopes for each of these antibodies, we have data to suggest that they have different epitopes and therefore will recognise a different sequence of amino acids within the protein. This is as far as we are aware, the only difference between these two antibodies.

    Read More

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