GITR/CD357 Rat mAb (Biotin) - Primary antibody, specific to GITR/CD357, Rat IgG2b

Cat. No.: Ab190289

GRADE & PURITY ExactAb™, Validated, 0.5 mg/mL

Application

Flow Cytometry

Host species

Rat

Reactive species

Mouse

Isotype

Rat IgG2b

Clone Number

DTA-1

Conjugation

Biotin

Overview

Rat anti Mouse GITR/CD357 Antibody, Monoclonal (DTA-1), could be used for Flow

Specifications

Product Name

GITR/CD357 Rat mAb (Biotin) - Primary antibody, specific to GITR/CD357, Rat IgG2b

Synonyms

Gitr | Activation-inducible TNFR family receptor | AITR | AITRTNF receptor superfamily activation-inducible protein | CD357 antigen | CD357 | GITR-D | GITRtumor necrosis factor receptor superfamily member 18 | Glucocorticoid-induced TNFR-related protein |

Specifications & Purity

ExactAb™, Validated, 0.5 mg/mL

Biochemical and Physiological Mechanisms

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.

Host species

Rat

Specificity

GITR/CD357

Immunogen

Mouse CD25+ CD4+ T cells

Positive Control

Flow: C57BL/6 mouse splenocytes.

Conjugation

Biotin

Grade

ExactAb™, Validated

Isotype

Rat IgG2b

Light Chain Type

lambda

Purification Method

Protein G purified

Concentration

0.5 mg/mL

Product Line

Biological Products

Storage and Shipping

Storage

Store at 2-8°C,Protected from light,Do not freeze

Shipped In

Wet ice,Do not freeze

Stability And Storage

The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.

Images

Flow Cytometry: GITR/CD357 Rat mAb (Biotin) (Ab190289)

GITR/CD357 Rat mAb (Biotin) (Ab190289) - Flow Cytometry
Two-color flow cytometric analysis of GITR/CD357 expression on mouse splenocytes. C57BL/6 mouse spleen cells were stained with CD25 Rat mAb (APC) (Ab179012) and either Rat IgG Isotype Control Antibody (Biotin) (Ab180310; Left panel) or GITR/CD357 Rat mAb (Biotin) (Ab190289; Right panel) at 1 μg/mL. Then incubated with Recombinant Streptavidin Protein (R-PE) (rp156150) at 0.1μg/test for 30 minutes at 4°C in the dark. Total viable cells were used for analysis.

Application

Application

Dilution info

Flow Cytometry

≤0.3 μg/test, the suggested use of this reagent is ≤0.3 μg per million cells in 100 µL staining volume

Associated Targets(Human)

TNFRSF18 Tbio Tumor necrosis factor receptor superfamily member 18 (0 Activities)

Discover Target: TNFRSF18

Activity Type	Activity Value -log(M)	Mechanism of Action	Activity Reference	Publications (PubMed IDs)

Mechanisms of Action

Application Protocols

Tested applications, recommended dilutions, incubation times, species reactivity, and positive controls are not specified for this item; refer to the CoA/Spec Sheet for authoritative usage guidance.

General recommendations (for planning only; not item-specific):

• Start with a titration matrix (e.g., 0.05–5 μg/mL for plate-based assays; 0.03–3 μg/test for flow cytometry) and optimize for signal-to-noise.
• Include appropriate controls: isotype, unstained, secondary-only (streptavidin-only), and known positive/negative samples.
• Maintain cold chain during setup; protect from light at all times due to biotinylated format possibly paired with fluorescent streptavidin.
• Document lot number, preparation date, and dilution to support reproducibility.

Because item-specific parameters are not provided here, finalize protocols based on your matrix, instrument, and detection system, and confirm with the CoA/Spec Sheet once available.

Biological Roles

Target background (general literature; not item-specific claims):

• GITR (Glucocorticoid-Induced TNFR-related protein), also known as CD357 and TNFRSF18, is a type I transmembrane receptor of the TNF receptor superfamily. It is expressed on T cells (notably Tregs and activated effector T cells), NK cells, and subsets of other immune cells.
• Ligand: GITRL (TNFSF18) expressed on antigen-presenting cells and some endothelial/epithelial cells.
• Signaling: Engagement of GITR can modulate NF-κB and MAPK pathways through TRAF adaptor interactions, affecting T-cell activation, proliferation, survival, and cytokine production.
• Immunology context: GITR is widely studied in immune regulation, particularly for its roles in modulating Treg function and costimulation of effector T cells. It is investigated in vaccination, tolerance, and tumor immunology models.

Research utility of anti-GITR antibodies (general):

• Phenotyping and quantifying GITR expression on immune subsets via flow cytometry.
• Functional studies involving receptor engagement/blockade or receptor occupancy measurements.
• Immunohistochemical localization of GITR-positive cells in tissues.

Caveats:

• Species orthologs may differ in sequence and expression; confirm species reactivity for the specific clone before use. Species reactivity for this SKU is not specified.
• Functional effects (agonism/antagonism) vary by clone and epitope; do not assume activity without validation data.

Reminder: This product is for research use only (from Product Data). No clinical or diagnostic use is implied.

Buffer Applications

No buffer composition is specified for this item; refer to the CoA/Spec Sheet. The guidance below is general best practice for handling biotinylated monoclonal antibodies.

• Working buffers (general):
  • PBS or TBS, pH 7.2–7.5, with 0.1–1% BSA or casein as blocker can reduce nonspecific binding in ELISA/flow/IHC.
  • Avoid free biotin in sample or diluent when a streptavidin readout is used; serum and tissue extracts can contain biotin—consider biotin blocking or preclearing steps.
  • For IHC, include appropriate detergent (e.g., 0.05–0.3% Tween-20) and serum/protein blockers as needed.
• Endogenous biotin (IHC/ICC): Use avidin/biotin blocking kits prior to applying biotinylated antibodies on tissues rich in endogenous biotin (e.g., liver, kidney).
• Storage buffer adjustments: Do not alter the supplied buffer unless necessary and validated; small pH or salt changes can affect stability and binding. For this item, supplied buffer is not specified—consult the CoA.
• Filtration/sterility: If sterility is required, use low-protein-binding 0.22 μm filters and validate post-filtration binding.

Note: If the product is supplied with preservatives (e.g., sodium azide) or protein carriers, these may influence compatibility with live cells or downstream conjugations. Presence/absence is not specified here.

Green Alternatives

While “green alternatives” typically address organic solvents and reagents, sustainability considerations also apply to antibody use and supply chains.

• Cold-chain efficiency: This item ships on wet ice and is stored at 2–8°C (do not freeze). Consolidate orders and coordinate deliveries to minimize shipments and associated packaging waste. Reuse or recycle cold packs and insulated shippers where permitted.
• Waste reduction: Optimize titrations to reduce reagent consumption. Aliquot responsibly to minimize spoilage; avoid unnecessary single-use plastics by selecting appropriately sized tubes and plates.
• Preservatives: If your workflow allows, prefer azide-free formulations for downstream live-cell work to avoid hazardous waste streams; however, the presence/absence of azide in this item is not specified—verify on the CoA/SDS.
• Process alternatives: For high-sensitivity detection, biotin–streptavidin amplification can allow reduced antibody usage compared to directly labeled fluorophores, lowering upstream manufacturing footprint per assay (general consideration).

Comparison (general):

• Biotinylated primary + streptavidin reporter: High sensitivity, modular; may require biotin blocking in tissues and careful buffer control.
• Directly labeled primary: Simpler workflow, reduced steps; typically lower amplification and potentially higher per-assay antibody consumption.

Note: No formulation specifics are provided for this SKU; implement institutional sustainability practices consistent with your assay needs.

Pharmaceutical Uses

This product is designated “For research use only” and is not intended for human or veterinary use, clinical diagnostics, or as a drug substance or excipient.

Formulation/manufacturing context (general information; not item-specific):

• Biotinylated antibodies are employed in analytical development and process monitoring assays (e.g., ELISA, ligand binding assays) to characterize biologics in preclinical research.
• In bioprocess analytics, biotin–streptavidin capture can enable high-specificity pull-downs for target quantitation or impurity profiling.

Regulatory note:

• No pharmacopeial monograph or excipient status is claimed or implied for this SKU. Any use in regulated environments requires independent qualification and documentation via the CoA, SDS, and method validation.

Item-specific details such as formulation grade for GMP/GLP, endotoxin level, or viral safety are not specified; consult Aladdin Scientific for documentation and suitability assessments if your application has compliance requirements.

Physical Properties

This product is a biotin-conjugated rat monoclonal antibody. Conventional small-molecule physical constants (BP, MP, logP, refractive index) are not applicable to proteins and are not provided for this item.

Item-specific properties:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer composition, preservative, and degree of labeling (DOL): Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item.

General/literature information for context (not item-specific):

• Typical IgG molecular mass is ~150 kDa (heavy–heavy–light–light tetramer) with glycosylation heterogeneity.
• Biotin-streptavidin binding is extraordinarily tight (Kd ~10^−14–10^−15 M, literature), enabling sensitive detection and robust capture.
• Solubility: Antibodies are water-soluble macromolecules formulated in aqueous buffers (e.g., PBS, TBS) with stabilizers; avoid extremes of pH and high ionic detergents unless validated (literature).
• Stability: Antibodies are sensitive to freeze–thaw and heat; maintain cold chain and protect from light for biotin conjugates (literature).

Practical notes:

• Do not infer buffer, pH, or additives for this specific item—use the CoA/Spec Sheet for authoritative formulation details.
• For assays requiring defined biotin density (e.g., multimer formation), verify the item’s DOL on the CoA if reported.

Quality & Grades

• Item-specific grade: ExactAb™, Validated (from Product Data).

What this generally implies (explanation; not a specification):

• ExactAb™: Denotes a controlled, reagent-grade monoclonal antibody offering consistent lot-to-lot performance under standardized manufacturing and QC practices. Exact meaning and criteria are brand-defined; consult the product CoA/Spec Sheet for the specific QC attributes included for this SKU.
• Validated: Indicates the antibody has undergone application-relevant performance checks. Because the exact tested applications, species reactivity, and acceptance criteria are not specified here, please refer to the CoA/Spec Sheet for the definitive validation scope (e.g., ELISA, flow cytometry, IHC), if provided.

What is not specified for this item:

• Clone ID, isotype/subclass, light chain type.
• Purity metrics (e.g., % monomer by SEC, residual aggregates), endotoxin level, binding affinity/avidity, and degree of labeling (DOL).
• Preservatives/stabilizers and formulation buffer.

Practical QA recommendations:

• When establishing assays, qualify the received lot in your workflow (e.g., titration series and known-positive/negative controls).
• Record lot number and retain a reference aliquot to support reproducibility.
• For regulated workflows, request or download the CoA, SDS, and any validation summaries to document fitness-for-purpose.

Reaction & Applications

Chemical reaction conditions are not applicable to antibody reagents. Instead, biotinylated antibodies are commonly used in biotin–streptavidin affinity and detection systems. Applications below are provided as general/literature guidance and are not item-specific claims.

• ELISA and plate-based assays: Use as capture or detection antibody in streptavidin-HRP/AP systems. Biotinylation enables flexible pairing with various streptavidin conjugates (enzymes, fluorophores).
• Flow cytometry: Detect cell-surface GITR/CD357 when combined with fluorophore-conjugated streptavidin. Benefits include signal amplification via multivalent streptavidin.
• Immunohistochemistry (IHC/ICC): Indirect detection on fixed tissues/cells with streptavidin–enzyme or –fluorophore detection; ensure endogenous biotin is blocked if tissue has high biotin levels.
• Western blot: Less common for cell-surface receptors without denaturation-resistant epitopes, but biotin-streptavidin systems can enhance sensitivity.
• Magnetic/affinity capture: Immobilize via streptavidin-coated beads or surfaces to enrich GITR-positive cells or immunoprecipitate receptor complexes.

Practical tips (general):

• Perform antibody titration to optimize signal-to-noise; biotinylated formats often require lower primary concentrations due to amplified detection.
• Include controls: isotype control, secondary-only (streptavidin-only) control, and known positive/negative biological samples.
• Confirm that sample diluents are devoid of free biotin; dialyze or desalt lysates if necessary.

Note: Tested applications, recommended dilutions, species reactivity, and clone/epitope for this item are not specified; consult the CoA/Spec Sheet.

Reaction Conditions

Not applicable to chemical synthesis. For assay workflows using biotinylated antibodies, the relevant “conditions” are biological/analytical rather than chemical.

General/literature guidance (not item-specific):

• ELISA: Coat plates with capture agent overnight at 4°C or 1–2 h at 37°C; block; apply biotinylated antibody at an empirically determined dilution; detect with streptavidin–HRP and TMB substrate.
• Flow cytometry: Incubate cells with biotinylated primary on ice or at 4°C for 20–30 min to minimize internalization; wash; develop with fluorophore-conjugated streptavidin; keep buffers cold and free of biotin.
• IHC/ICC: Optimize antigen retrieval, blocking, and detergent; apply biotinylated primary; detect with streptavidin–enzyme/fluorophore systems; include endogenous biotin block where needed.

These are general practices. Tested applications, recommended dilutions, incubation times, and temperatures for this specific SKU are not provided—consult the CoA/Spec Sheet and perform empirical optimization.

Safety & Handling

• GHS/CLP classification, signal word, hazard statements, and pictograms: Not specified for this item; consult the SDS for authoritative safety information.
• Storage conditions (Product Data): Store at 2–8°C, protected from light. Do not freeze. Shipped on wet ice (do not freeze in transit).
• General laboratory PPE: Lab coat, gloves (nitrile or equivalent), and safety glasses are recommended when handling any biological reagent.
• Handling guidance (general):
  • Avoid microbial contamination; work aseptically if repeated access is expected.
  • Minimize light exposure to preserve the integrity of the biotin conjugate and any light-sensitive excipients (if present).
  • Avoid repeated freeze–thaw cycles; this item is explicitly “Do not freeze.”
  • Do not mix with oxidizing biotin-blocking agents when subsequent streptavidin/avidin steps are required.
• Incompatibilities (general): Strong oxidants, extremes of pH, and denaturing surfactants or solvents can compromise antibody structure and antigen binding.
• First-aid (general overview; defer to SDS):
  • Skin/eye contact: Rinse with water; remove contaminated clothing; seek medical advice if irritation persists.
  • Ingestion/inhalation: Rinse mouth, move to fresh air, and obtain medical attention if unwell.
• Waste disposal: Follow institutional biosafety procedures for proteinaceous laboratory waste; dispose according to local regulations.

Note: Absence of sodium azide, glycerol, or other preservatives is not implied—formulation details are not specified for this item and must be verified on the CoA/SDS.

Solvent Selection

Not a solvent. This product is an aqueous protein reagent (biotinylated rat monoclonal antibody). Conventional organic solvent selection does not apply.

General guidance (literature/practice) for buffer environments:

• Typical compatible buffers for antibodies include PBS or TBS at near-neutral pH (7.2–7.5). Avoid unvalidated high concentrations of denaturants, chaotropes, or organic cosolvents that can disrupt tertiary structure and antigen binding.
• For biotin–streptavidin workflows, ensure sample buffers are free of free biotin and strong biotin competitors. Include carrier proteins (e.g., BSA) only if they do not interfere with downstream avidin/streptavidin steps.
• Detergents: If needed for tissue/permeabilization workflows, use mild, nonionic detergents at validated concentrations; verify compatibility with your assay.

For this specific item, buffer composition and additives are not specified; refer to the CoA/Spec Sheet before adjusting matrices.

Storage & Reconstitution

• Storage (Product Data): Store at 2–8°C. Protect from light. Do not freeze. Shipped on wet ice (do not freeze during transit).
• Format: Not specified (liquid vs. lyophilized) for this item; refer to the CoA/Spec Sheet and product label.

Handling and aliquoting (general best practice):

• If supplied as a liquid, mix gently by inversion; avoid vigorous vortexing that may induce aggregation or foaming.
• If aliquoting, use low-protein-binding plastics and prechilled equipment. Avoid multiple freeze–thaw cycles; this item is explicitly “Do not freeze.”
• Maintain sterile technique to prevent contamination during repeated use.

Reconstitution (only if lyophilized; not specified here):

• Follow label instructions. Typical practice is reconstitution with sterile PBS or water to the recommended concentration, then gentle mixing and brief equilibration on ice.

Light protection:

• Keep in amber vials or wrap tubes/plates in foil during storage and incubations, especially when paired with fluorescent streptavidin.

Stability and expiration:

• Shelf life and on-bench/in-use stability are not specified for this item; consult the CoA/Spec Sheet and observe manufacturer’s expiration dating on the label.

Note: Do not infer presence/absence of preservatives or carriers; confirm on the CoA/SDS before deciding on storage strategy.

Structure & Identity

GITR/CD357 Rat mAb (Biotin) is a rat-derived monoclonal antibody directed against the cell-surface receptor GITR (also known as CD357; TNFRSF18). It is supplied as a biotin-conjugated immunoreagent for streptavidin/avidin-based detection and capture workflows.

• Item-specific identifiers (from Product Data):
  • SKU: Ab190289
  • Grade/Purity: ExactAb™, Validated
  • Conjugate: Biotin
  • Target name: GITR/CD357 (TNFRSF18)
  • Host: Rat monoclonal antibody
• Small-molecule identifiers (CAS, CID, InChIKey, SMILES): Not applicable to antibodies; not specified for this item.
• Molecular formula and exact molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Structural features (general, literature):
  • Typical IgG monoclonal architecture comprising two heavy chains (~50 kDa each) and two light chains (~25 kDa each) with inter- and intra-chain disulfides, forming a Y-shaped 2D topology with antigen-binding Fab arms and an Fc domain.
  • Biotin conjugation generally occurs via NHS ester chemistry to accessible lysine ε-amines on the antibody surface, yielding multiple biotin moieties for high-affinity interaction with streptavidin/avidin (general literature).
  • Antigen: GITR/CD357 is a type I transmembrane glycoprotein receptor of the TNF receptor superfamily (general literature).

Notes:

• No clone ID, isotype, light-chain type, or degree of labeling (DOL) are specified for this item; consult CoA/Spec Sheet if needed.
• All structure descriptions beyond item-specific identifiers are general antibody conventions (literature).

Synthetic Utility

Not applicable. This product is a biotin-conjugated monoclonal antibody intended for immunodetection, capture, and related bioanalytical workflows, not for use as a synthetic reagent or building block.

General note (for context only): Antibodies can be components in chemoenzymatic conjugations or surface functionalization workflows (e.g., immobilization on streptavidin-coated materials via biotin), but these are affinity-driven assemblies rather than chemical synthesis in the conventional sense.

Target Specificity

Item-specific details available:

• Target name: GITR/CD357 (TNFRSF18) — as stated in the product name.
• Antibody type: Rat monoclonal antibody.
• Conjugate: Biotin.

Not specified for this item (refer to CoA/Spec Sheet if required):

• Clone ID and epitope/region recognized.
• Isotype/subclass and light-chain type.
• Validated species reactivity and cross-reactivity profile.
• Functional characteristics (e.g., neutralizing, agonistic, depleting).

General background (for context only; not item-specific claims): GITR/CD357 is a TNF receptor family member expressed on T cells and other immune subsets. Specificity parameters (affinity, epitope binning, off-target binding) are clone-dependent and must be verified for the particular antibody lot.