Recombinant
RabMAb

Recombinant Anti-KIR2DL1 + KIR2DL2 antibody [EPR22492-2] (ab224696)

Overview

  • Product name

    Anti-KIR2DL1 + KIR2DL2 antibody [EPR22492-2]
  • Description

    Rabbit monoclonal [EPR22492-2] to KIR2DL1 + KIR2DL2
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human KIR2DL1 aa 1-250. The exact sequence is proprietary.
    Database link: P43626

  • Positive control

    • WB: His-tagged human KIR2DL1 recombinant protein (aa1-245); His and LIF-tagged human KIR2DL2 recombinant protein (aa26-221). IHC-P: Human endometrium tissue. ICC/IF: NK-92 cells (treated with 5-aza-2-deoxycytidine (2µM) for 48h). Flow Cyt: Human PBMCs. IP: NK-92 (treated with 2µM 5-aza-2-deoxycytidine for 48h) whole cell 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 ab224696 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
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.
Flow Cyt 1/60.
IP 1/30.
  • Application notes
    Is unsuitable for WB.
  • Target

    Images

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NK-92 (human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells (+/- treatment with 5-aza-2-deoxycytidine(2µM) for 48h) labeling KIR2DL1 + KIR2DL2 with ab224696 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in NK-92 cells treated with 5-aza-2-deoxycytidine(2uM) for 48h. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling KIR2DL1 + KIR2DL2 with ab224696 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in natural killer (NK) cells in human endometrium (PMID: 7749980) is observed. Counterstained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • KIR2DL1 + KIR2DL2 were immunoprecipitated from 0.35 mg of NK-92 (human peripheral blood malignant non-Hodgkin's lymphoma cell line) (treated with 2μM 5-aza-2-deoxycytidine for 48h) whole cell lysate with ab224696 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224696 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

      Lane 1: NK-92 (treated as above) whole cell lysate 10 μg (Input).
      Lane 2: ab224696 IP in NK-92 (treated as above) whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224696 in NK-92 (treated as above) whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 100 seconds.

      This blot was developed using a higher sensitivity ECL substrate.

    • Flow cytometric analysis of human PBMCs (peripheral blood mononuclear cells) labeling KIR2DL1 + KIR2DL2 with ab224696 at 1/60 dilution (Right) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).

      Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

      Cells were stained with rabbit IgG (Left) or ab224696 (Right). Then stained with anti-CD56 conjugated to BV421.

      Gated on viable cells.

       

    • All lanes : Anti-KIR2DL1 + KIR2DL2 antibody [EPR22492-2] (ab224696) at 1/1000 dilution

      Lane 1 : His-tagged human KIR2DL1 recombinant protein (aa1-245) 20 ng
      Lane 2 : His and LIF-tagged human KIR2DL2 recombinant protein (aa26-221) 20 ng
      Lane 3 : His-tagged human KIR2DL3 recombinant protein (aa1-245) 20 ng
      Lane 4 : His-tagged human KIR2DS1 recombinant protein (aa22-221) 20 ng
      Lane 5 : His-tagged human KIR2DS2 recombinant protein (aa22-221) 20 ng
      Lane 6 : His-tagged human KIR2DS3 recombinant protein (aa22-221) 20 ng
      Lane 7 : His-tagged human KIR2DS4 recombinant protein (aa22-221) 20 ng
      Lane 8 : His-tagged human KIR2DS5 recombinant protein (aa22-221) 20 ng

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

      Predicted band size: 39 kDa


      Exposure time: 26 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    References

    ab224696 has not yet been referenced specifically in any publications.

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