Overview

  • Product name

    Anti-PIGK antibody [EPR17843]
    See all PIGK primary antibodies
  • Description

    Rabbit monoclonal [EPR17843] to PIGK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PIGK aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: Q92643

  • Positive control

    • WB: HT1080, HEK-293 and C6 cell lysates; Human pancreas and fetal liver lysates; Mouse brain, mouse heart, mouse kidney, mouse spleen, rat brain and rat kidney lysates. ICC/IF: A549 and Jurkat cells. Flow Cyt: HEK-293 cells. IP: HEK-293 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17843
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab201693 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
ICC/IF 1/500.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP 1/30.
WB 1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).

Target

  • Relevance

    PIGK is a member of the cysteine protease family C13 that is involved in glycosylphosphatidylinositol (GPI)-anchor biosynthesis. The GPI anchor is a glycolipid found on many blood cells and serves to anchor proteins to the cell surface. This protein is a member of the multisubunit enzyme, GPI transamidase and is thought to be its enzymatic component. GPI transamidase mediates GPI anchoring in the endoplasmic reticulum, by catalyzing the transfer of fully assembled GPI units to proteins.
  • Cellular localization

    Endoplasmic reticulum, Endoplasmic reticulum membrane; type I single pass membrane protein
  • Database links

  • Alternative names

    • GPI anchor transamidase antibody
    • GPI transamidase antibody
    • GPI8 antibody
    • Phosphatidylinositol glycan biosynthesis class K protein antibody
    • Phosphatidylinositol glycan class K antibody
    • PIG K antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PIGK with ab201693 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic staining on Jurkat cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab201693 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling PIGK with ab201693 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic staining on A549 cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab201693 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • All lanes : Anti-PIGK antibody [EPR17843] (ab201693) at 1/2000 dilution

    Lane 1 : HT1080 (Human fibrosarcoma cells) cell lysate
    Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PIGK antibody [EPR17843] (ab201693) at 1/2000 dilution

    Lane 1 : Human pancreas lysate
    Lane 2 : Human fetal liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PIGK antibody [EPR17843] (ab201693) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : C6 (Rat glial tumor cells) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HEK-293 (Human epithelial cells from embryonic kidney) cells labeling PIGK with ab201693 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • PIGK was immunoprecipitated from 1mg of HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab201693 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab201693 at 1/2000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

    Lane 1: HEK293 whole cell lysate10 µg (Input).

    Lane 2: ab201693 IP in HEK293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201693 in HEK293 whole cell lysate.

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

    Exposure time: 5 seconds.

References

ab201693 has not yet been referenced specifically in any publications.

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