Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139]

6 product images

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

  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

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

  Lane 1: Human spleen tissue lysate at 60 µg

  Lane 2: Human tonsil tissue lysate at 60 µg

  Secondary

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

  Observed band size: 60-80 kDa

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  Flow Cytometry - Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control. Goat anti rabbit IgG (Alexa Fluor^® 647, Goat Anti-Rabbit IgG H&L (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.

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

  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

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

  Lane 1: His-tagged human KIR3DL1 recombinant protein at 0.01 µg

  Lane 2: His-tagged human KIR3DL3 recombinant protein at 0.01 µg

  Lane 3: His-tagged human KIR2DS4 recombinant protein at 0.01 µg

  Lane 4: LIF/His-tagged human KIR2DS2 recombinant protein at 0.01 µg

  Secondary

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

  Observed band size: 33-87 kDa

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

  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

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

  Lanes 1, 3 and 5: 293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T transfected with human KIR3DL1 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 4: 293T transfected with human KIR3DL2 expression vector containing a myc-His-tag®, whole cell lysate 20 at 20 µg

  Lane 6: 293T transfected with human KIR3DS1 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Secondary

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

  Observed band size: 45-70 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/500 dilution.

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  Immunoprecipitation - Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

  All lanes: Immunoprecipitation - Anti-KIR3DL1 + KIR2DS4 + KIR2DS2 + KIR3DS1 antibody [EPR24357-139] (ab259808)

  Observed band size: 60-80 kDa