Recombinant

Recombinant Anti-GITR antibody [CAL52] - BSA and Azide free (ab251600)

Overview

  • Product name
    Anti-GITR antibody [CAL52] - BSA and Azide free
    See all GITR primary antibodies
  • Description
    Rabbit monoclonal [CAL52] to GITR - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IP, Flow Cytmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human GITR aa 150-250.
    Database link: Q9Y5U5

  • Positive control
    • IHC-P: Human tonsil tissue. ICC/IF: HEK-293T cells. IP: HEK-293T and Hut-78 whole cell lysate. Flow: Human peripheral blood mononuclear cells.
  • General notes

    ab251600 is a PBS-only buffer format of ab237713. Please refer to ab237713 for recommended dilutions, protocols, and image data.

     

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

     This product was previously labelled as TNFRSF18

     

Properties

Applications

Our Abpromise guarantee covers the use of ab251600 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 Use at an assay dependent concentration.

Boil tissue section in TRIS EDTA buffer for 24 min followed by cooling at room temperature for 30-45 min. Primary antibody incubation for 75 minutes at room temperature.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function
      Receptor for TNFSF18. Seems to be involved in interactions between activated T-lymphocytes and endothelial cells and in the regulation of T-cell receptor-mediated cell death. Mediated NF-kappa-B activation via the TRAF2/NIK pathway.
    • Tissue specificity
      Expressed in lymph node, peripheral blood leukocytes and weakly in spleen.
    • Sequence similarities
      Contains 3 TNFR-Cys repeats.
    • Cellular localization
      Secreted and Cell membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • Activation inducible TNFR family receptor antibody
      • Activation-inducible TNFR family receptor antibody
      • AITR antibody
      • CD357 antibody
      • GITR antibody
      • GITR D antibody
      • GITR-D antibody
      • GITRD antibody
      • Glucocorticoid induced TNFR related protein antibody
      • glucocorticoid-induced tnf receptor ligand antibody
      • glucocorticoid-induced tnf receptors ligand antibody
      • Glucocorticoid-induced TNFR-related protein antibody
      • TNF receptor superfamily activation inducible protein antibody
      • TNFRSF 18 antibody
      • TNFRSF18 antibody
      • TNR18_HUMAN antibody
      • Tumor necrosis factor receptor superfamily member 18 antibody
      • Tumor necrosis factor receptor superfamily member 18 precursor antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling GITR with ab237713 at 1/4000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human tonsil is observed. Counter stained with Hematoxylin. The section was incubated with ab237713 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713).

    • Formalin-fixed, paraffin-embedded human tonsil tissue stained for TNFSF18 using ab237713 at 0.25 μg/ml in immunohistochemical analysis.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713).

    • 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labeling GITR with ab237713 at 1/50 dilution followed by a AlexaFluor®594 Goat anti-Rabbit secondary (ab150080) at a 1/500 dilution (Green). The nuclear counterstain was DAPI (Blue). Confocal image showing Positive staining in HEK-293T cells transfected with a GFP-tagged GITR expression construct.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor®594 Goat anti-Rabbit secondary (ab150080) at a 1/500 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713). 

    • Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) treated with 10μg/ml PHA for 48h, labeling GITR with ab237713 at 1/500 dilution. The secondary antibody was a Goat anti rabbit IgG (Alexa Fluor® 488, ab150097) at 1/500 dilution. Cells were surface stained with anti-CD25 conjugated to BV421. Then fixed with 2% PFA followed by intracellular staining rabbit IgG (Left) or ab237713 (Right). The isotype control used was a Rabbit monoclonal IgG (ab172730, Left).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713).

    • GITR was immunoprecipitated from 0.35 mg Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate using ab237713 at 1/30 dilution. western blot was performed on the immunoprecipitate using ab237713 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

      Lane 1: Hut-78 whole cell lysate 10 μg (input)
      Lane 2: ab237713 IP in Hut-78 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237713 in Hut-78 whole cell lysate.

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

      Exposure time: 15 seconds.

      This blot was developed using a higher sensitivity ECL substrate.

      Dimerized GITR was also observed at 52kDa

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713).

    • GITR was immunoprecipitated from 0.35 mg HEK-293T (Human embryonic kidney epithelial cell) transfected with GFP-tagged GITR overexpression vector whole cell lysate using ab237713 at 1/30 dilution. western blot was performed on the immunoprecipitate using ab237713 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

      Lane 1: Hut-78 whole cell lysate 10 μg (input)
      Lane 2: ab237713 IP in Hut-78 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237713 in 293T transfected with GFP-tagged GITR overexpression vector whole cell lysate

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

      Exposure time: 3 minutes.

      This blot was developed using a higher sensitivity ECL substrate.

      Dimerized GITR was also observed at 52kDa

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237713).

    References

    ab251600 has not yet been referenced specifically in any publications.

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