Recombinant
RabMAb

Recombinant Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

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Overview

  • Product name

    Anti-SMARCD2 antibody [EPR20860-251]
    See all SMARCD2 primary antibodies

  • Description

    Rabbit monoclonal [EPR20860-251] to SMARCD2

  • Host species

    Rabbit

  • Specificity

    IHC is recommended only for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human SMARCD2 aa 300-400. The exact sequence is proprietary.
    Database link: Q99JR8

  • Positive control

    • WB: Neuro-2a cells transfected with scrambled siRNA control, whole cell lysate; mouse P70 thymus, white adipose, brown adipose and liver lysate; Beta-TC-6 whole cell lysate; Rat thymus lysate; Jurkat and HeLa whole cell lysate. IHC-P: Mouse colon tissue; Rat testis tissue. Flow cyt: HeLa cells. IP: HeLa whole cell lysate; Mouse P70 thymus lysate.

  • 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 ab220164 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. Detects a band of approximately 59 kDa (predicted molecular weight: 59 kDa).
IHC-P 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/600.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Relevance

      SMARCD2 is a member of the SWI/SNF family of proteins which are thought to regulate transcription of certain genes by altering the chromatin structure around those genes. It is part of the large ATP-dependent chromatin remodeling complex SNF/SWI.

    • Cellular localization

      Nuclear

    • Database links

      see all

    • Alternative names

      • 60 kDa BRG 1/Brm associated factor subunit B antibody
      • 60 kDa BRG1/Brm associated factor subunit B antibody
      • BAF60B antibody
      • BRG1 associated factor 60B antibody
      • Chromatin remodeling complex BAF60B subunit antibody
      • CRACD 2 antibody
      • CRACD2 antibody
      • Mammalian chromatin remodeling complex BRG1 associated factor 60B antibody
      • PRO2451 antibody
      • Rsc6p antibody
      • SMARCD 2 antibody
      • SWI/SNF complex 60 kDa subunit B antibody
      • SWI/SNF related matrix associated actin dependent regulator of chromatin d2 antibody
      • SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily d member 2 antibody
      • Swp73 like protein antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

      Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling SMARCD2 with ab220164 at 1/200 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat testis (PMID: 20148946, 28303890, 25622893). The section was incubated with ab220164 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunoprecipitation - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Immunoprecipitation - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

      SMARCD2 was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab220164 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220164 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

      Lane 1: HeLa whole cell lysate 10 µg (Input).
      Lane 2: ab220164 IP in HeLa whole cell lysate .
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220164 in HeLa whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 40 seconds.

    • Flow Cytometry - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Flow Cytometry - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling SMARCD2 with ab220164 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

      Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

      Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling SMARCD2 with ab220164 at 1/200 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse colon (PMID: 20148946, 28303890, 25622893). The section was incubated with ab220164 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunoprecipitation - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Immunoprecipitation - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)

      SMARCD2 was immunoprecipitated from 0.35 mg mouse P70 thymus lysate with ab220164 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220164 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

      Lane 1: Mouse P70 thymus lysate 10 µg (Input).
      Lane 2: ab220164 IP in mouse P70 thymus lysate .
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220164 in mouse P70 thymus lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 seconds.

    • Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      All lanes : Anti-SMARCD2 antibody [EPR20860-251] (ab220164) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 3 : Rat thymus lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 59 kDa
      Observed band size: 59 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lane 1: 59 secs; Lane 2: 81 secs; Lane 3: 26 secs.

    • Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      All lanes : Anti-SMARCD2 antibody [EPR20860-251] (ab220164) at 1/1000 dilution

      Lane 1 : Mouse P70 liver lysate
      Lane 2 : Mouse P70 thymus lysate
      Lane 3 : Mouse P70 white adipose lysate
      Lane 4 : Mouse P70 brown adipose lysate
      Lane 5 : Beta-TC-6 (mouse pancreas insulinoma beta cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 59 kDa
      Observed band size: 59 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      Western blot - Anti-SMARCD2 antibody [EPR20860-251] (ab220164)
      All lanes : Anti-SMARCD2 antibody [EPR20860-251] (ab220164) at 1/1000 dilution

      Lane 1 : Neuro-2a (mouse neuroblastoma cell line) transfected with scrambled siRNA control whole cell lysate
      Lane 2 : SMARCD2 knockdown Neuro-2a whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 59 kDa
      Observed band size: 59 kDa


      Exposure time: 3 minutes


      Lysates were kindly provided by Dr Zhuoxian Meng.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    References

    ab220164 has not yet been referenced specifically in any publications.

    Publishing research using ab220164? Please let us know so that we can cite the reference in this datasheet.

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