Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

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 - BSA and Azide free
  • Suitable for: ICC, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-ARID1A antibody [EPR13501] - BSA and Azide free
    See all ARID1A primary antibodies

  • Description

    Rabbit monoclonal [EPR13501] to ARID1A - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • IHC-P: Human kidney tissue, Human adenocarcinoma of endometrium without ARID1A mutation. Flow Cyt: SH-SY5Y cells. ICC/IF: ARID1A wildtype and knockout HAP1 cells WB: HEK-293T and SH-SY5Y cell lysates.

  • General notes

    Ab217154 is the carrier-free version of ab182560. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab217154 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Western blot - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab182560).

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Flow Cytometry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).

  • Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    This IHC data was generated using the same anti-ARID1A antibody clone, EPR13501, in a different buffer formulation (cat# 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.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    Clone EPR13501 (ab217154) has been successfully conjugated by Abcam. This image was generated using Anti-ARID1A antibody [EPR13501] (Alexa Fluor® 647). Please refer to ab216304 for protocol details.

    ab216304 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 ab216304 at 1/500 dilution (shown in red) and ab195887 at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    Clone EPR13501 (ab217154) has been successfully conjugated by Abcam. This image was generated using Anti-ARID1A antibody [EPR13501] (Alexa Fluor® 488). Please refer to ab216112 for protocol details.

    ab216112 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 ab216112 at 1/500 dilution (shown in green) and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Immunocytochemistry - Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).

  • Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)
    Anti-ARID1A antibody [EPR13501] - BSA and Azide free (ab217154)

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