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RecombinantRabMAb

Recombinant Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139] (ab259808)

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Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Immunocytochemistry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Flow Cytometry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Immunoprecipitation - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
  • Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24357-139] to KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1
  • Suitable for: IP, ICC, WB, Flow Cyt
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647

Overview

  • Product name

    Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139]
    See all KIR3DL1+KIR2DS4+KIR2DS2+KIR3DS1 primary antibodies

  • Description

    Rabbit monoclonal [EPR24357-139] to KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC, WB, Flow Cytmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: Human spleen, His-tagged human KIR3DL1 recombinant protein, His-tagged human KIR2DS4 recombinant protein, 293T transfected with human KIR3DL1expression vector containing a myc-His-tag®, 293T transfected with human KIR3DS1 expression vector containing a myc-His-tag® whole lysates. ICC: 293T cells transfected with myc-tagged KIR3DL1 expression vector. Flow Cyt: Human peripheral blood mononuclear cells. IP: 293T transfected with human KIR3DL1expression vector containing a myc-His-tag®.

  • 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 ab259808 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
IP
1/30.
ICC
1/50.
WB
1/1000. Predicted molecular weight: 49 kDa.
Flow Cyt
1/500.
Notes
IP
1/30.
ICC
1/50.
WB
1/1000. Predicted molecular weight: 49 kDa.
Flow Cyt
1/500.
Application notes
Is unsuitable for IHC-P.

Target

Images

  • Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    All lanes : Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139] (ab259808) at 1/1000 dilution

    Lane 1 : Human spleen tissue lysate
    Lane 2 : Human tonsil tissue lysate

    Lysates/proteins at 60 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 49 kDa
    Observed band size: 60-80 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression pattern & observed MW are consistent with what has been described in the literature (PMID:17911614).

    Negative control: human tonsil (PMID:14734722)

    Exposure time: 3 minutes

     

  • Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    All lanes : Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139] (ab259808) at 1/1000 dilution

    Lane 1 : His-tagged human KIR3DL1 recombinant protein
    Lane 2 : His-tagged human KIR3DL3 recombinant protein
    Lane 3 : His-tagged human KIR2DS4 recombinant protein
    Lane 4 : LIF/His-tagged human KIR2DS2 recombinant protein

    Lysates/proteins at 0.01 µg per lane.

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

    Predicted band size: 49 kDa
    Observed band size: 33-87 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    This antibody has high cross-reactivity with human KIR2DS4, and low with human KIR2DS2. This antibody does not recognize human KIR3DL3.

    All of the rec proteins were made in-house. KIR3DL1 and KIR2DS2 were expressed from HEK-293 expression systems. KIR3DL3 and KIR2DS4 were expressed from E.coli expression systems.

    Exposure time: 15 seconds

  • Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Western blot - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    All lanes : Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139] (ab259808) at 1/1000 dilution

    Lanes 1 & 3 & 5 : 293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
    Lane 2 : 293T transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate
    Lane 4 : 293T transfected with human KIR3DL2 expression vector containing a myc-His-tag®, whole cell lysate 20
    Lane 6 : 293T transfected with human KIR3DS1 expression vector containing a myc-His-tag®, 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 49 kDa
    Observed band size: 45-70 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    This antibody can recognize human KIR3DS1 but can not recognize human KIR3DL2.

    Exposure time: Lane 1-4: 3 minutes Lane 5-6: 59 seconds 

  • Immunocytochemistry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Immunocytochemistry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney) cells labelling KIR3DL1+KIR3DS1+KIR2DS2+KIR2DS4 with ab259808 at 1/50 dilution (10.2 ug/ml), followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with myc-tagged KIR3DL1 expression vector is observed.

    Myc-Tag Mouse mAb (Alexa Fluor® 488) was used to counterstain myc-tag at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 dilution.

  • Flow Cytometry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Flow Cytometry - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)

    Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling KIR3DL1+KIR3DS1+KIR2DS2+KIR2DS4 with ab259808 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. Goat anti rabbit IgG (Alexa Fluor® 647, ab150079) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab259808. Then stained with anti-CD56 conjugated to BV421. Gated on viable cells.

  • Immunoprecipitation - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Immunoprecipitation - Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)

    KIR3DL1+KIR3DS1+KIR2DS2+KIR2DS4 was immunoprecipitated from 0.35 mg 293T transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate with ab259808 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259808 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: 293T (human embryonic kidney) transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate 10 ug

    Lane 2: ab259808 IP in 293T transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259808 in 293T transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

  • Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)
    Anti-KIR3DL1 antibody [EPR24357-139] (ab259808)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab259808? Please let us know so that we can cite the reference in this datasheet.

ab259808 has not yet been referenced specifically in any publications.

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