Recombinant Anti-GITR antibody [CAL52] - BSA and Azide free (ab251600)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAL52] to GITR - BSA and Azide free
- Suitable for: IHC-P, Flow Cyt (Intra), ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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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, Flow Cyt (Intra), ICC/IF, IPmore details
Unsuitable for: WB -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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.
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General notes
ab251600 is the carrier-free version of ab237713.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purity is greater than 99%. -
Clonality
Monoclonal -
Clone number
CAL52 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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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. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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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. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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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
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Database links
- Entrez Gene: 8784 Human
- Omim: 603905 Human
- SwissProt: Q9Y5U5 Human
- Unigene: 212680 Human
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Alternative names
- Activation inducible TNFR family receptor antibody
- Activation-inducible TNFR family receptor antibody
- AITR antibody
see all
Images
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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).
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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).
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Intracellular 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 GITRwith 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).
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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).
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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 lysateBlocking 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).
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.
Datasheets and documents
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Datasheet download
References (0)
ab251600 has not yet been referenced specifically in any publications.