Recombinant
RabMAb

Recombinant Anti-GNLY/Granulysin antibody [EPR22110-128] (ab223326)

Overview

  • Product name

    Anti-GNLY/Granulysin antibody [EPR22110-128]
    See all GNLY/Granulysin primary antibodies
  • Description

    Rabbit monoclonal [EPR22110-128] to GNLY/Granulysin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human GNLY/Granulysin aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P22749

  • Positive control

    • WB: His-tagged human GNLY/Granulysin recombinant protein (aa23-145) ; NK-92 whole cell lysate. IP: NK-92 whole cell lysate. ICC/IF: NK-92 cells. Flow cyt: PBMCs.
  • General notes

     This product was previously labelled as GNLY

     

    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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22110-128
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab223326 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 14, 13 kDa (predicted molecular weight: 16 kDa).
ICC/IF 1/1000.
IP 1/30.
Flow Cyt 1/600.

Target

  • Function

    Antimicrobial protein that kills intracellular pathogens. Active against a broad range of microbes, including Gram-positive and Gram-negative bacteria, fungi, and parasites. Kills Mycobacterium tuberculosis.
  • Tissue specificity

    Expressed in natural killer and T-cells.
  • Sequence similarities

    Contains 1 saposin B-type domain.
  • Post-translational
    modifications

    A 9 kDa form is produced by proteolytic processing of a 15 kDa protein.
  • Cellular localization

    Secreted. Located in the cytotoxic granules of T-cells, which are released upon antigen stimulation.
  • Information by UniProt
  • Database links

  • Alternative names

    • 519 antibody
    • D2S69E antibody
    • GNLY antibody
    • GNLY_HUMAN antibody
    • Granulysin antibody
    • Granulysin precursor antibody
    • LAG 2 antibody
    • LAG2 antibody
    • Lymphocyte activation gene 2 antibody
    • Lymphokine LAG 2 antibody
    • Lymphokine LAG-2 antibody
    • NKG 5 antibody
    • NKG5 antibody
    • Protein NKG5 antibody
    • T cell activation protein 519 antibody
    • T lymphocyte activation gene 519 antibody
    • T-cell activation protein 519 antibody
    • TLA519 antibody
    see all

Images

  • All lanes : Anti-GNLY/Granulysin antibody [EPR22110-128] (ab223326) at 1/1000 dilution

    Lane 1 : NK-92 (human peripheral blood malignant non-Hodgkin's lymphoma cell line) whole cell lysate
    Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow) 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: 16 kDa
    Observed band size: 13,14 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

    The doublets likely represent the two isoforms of GNLY/Granulysin according to the UniProt Database.

    Negative control: K-562 cell line (PMID:2434598).

    This blot was developed using high sensitivity ECL substrate.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20-permeabilized PBMCs (human peripheral blood mononuclear cell) cells labeling GNLY/Granulysin with ab223326 at 1/600 (right) compared with a Isotype control details (ab172730) (left).  

    Human PBMCs were stained with PE-conjugated anti-human CD56.
    Cells were then fixed with 2% PFA for 10min and permeabilized with 0.1% Tween-20, followed by intracellular staining with rabbit IgG (ab172730, left) or ab223326, right).

  • GNLY/Granulysin was immunoprecipitated from 0.35 mg of NK-92 (human peripheral blood malignant non-Hodgkin's lymphoma cell line) whole cell lysate with ab223326 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223326 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: NK-92 whole cell lysate 10 μg (Input).
    Lane 2: ab223326 IP in NK-92 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223326 in NK-92 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 10 seconds.

    The doublets likely represent the two isoforms of GNLY/Granulysin according to the UniProt Database.

  • 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 labeling GNLY/Granulysin with ab223326 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in the NK-92 cell line is observed. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    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.

    Negative control: K-562 (PMID: 2434598).

  • Anti-GNLY/Granulysin antibody [EPR22110-128] (ab223326) at 1/1000 dilution + His-tagged human GNLY/Granulysin recombinant protein (aa23-145) at 0.01 µg

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

    Predicted band size: 16 kDa
    Observed band size: 14 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

References

ab223326 has not yet been referenced specifically in any publications.

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