GIT1 Antibody - Anticuerpo primario, specific to GIT1, Rabbit IgG

Cat. No.: Ab105406

CALIDAD & PUREZA Carrier Free, ExactAb™, Validated, 1.0 mg/mL

Application

IHC, WB

Host species

Conejo

Reactive species

consejo, Humano, Ratón

Isotype

Rabbit IgG

Conjugation

Unconjugated

Descripción general

Rabbit anti Human GIT1 Antibody, Polyclonal, could be used for WB, IHC and so on.
Application:
WB: 1/500-1/1000
IHC: 1/50-1/100
Protein Function：
GTPase-activating protein for the ADP ribosylation factor family. May serve as a scaffold to bring together molecules to form signaling modules controlling vesicle trafficking, adhesion and cytoskeletal organization. Increases the speed of cell migration, as well as the size and rate of formation of protrusions, possibly by targeting PAK1 to adhesions and the leading edge of lamellipodia. Sequesters inactive non-tyrosine-phosphorylated paxillin in cytoplasmic complexes.

Especificaciones

Product Name

GIT1 Antibody - Anticuerpo primario, specific to GIT1, Rabbit IgG

Sinónimos

ARF GAP GIT1 | ARF GTPase-activating protein GIT1 | CAT1 | CAT-1 | Cool-associated and tyrosine-phosphorylated protein 1 | G protein-coupled receptor kinase interacting ArfGAP 1 | G protein-coupled receptor kinase interactor 1 | G protein-coupled receptor

Especificaciones y pureza

ExactAb™, Validated, Carrier Free, 1.0 mg/mL

Especies hospedadoras

Conejo

Especificidad

GIT1

Inmunógeno

A synthetic peptide derived from human GIT1

Control positivo

WB: A549, HeLa, Hep G2, NIH/3T3 and C6 cell lysates. IHC: Human lung cancer tissue.

Conjugación

Unconjugated

Grado

Carrier Free, ExactAb™, Validated

Isotipo

Rabbit IgG

SDS-PAGE

150 kDa

Método de purificación

Immunogen affinity purified

Concentración

1.0 mg/mL

Product Line

Productos biológicos

Almacenamiento y envío

Forma

Liquid

Condiciones de almacenamiento de almacenamiento

Store at -20°C,Avoid repeated freezing and thawing

Enviado en

Ice chest + Ice pads

Estabilidad y almacenamiento

Store at 4°C short term (1-2 weeks). Store at -20°C long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Imágenes

Western Blot: GIT1 Antibody (Ab105406)

GIT1 Antibody (Ab105406) - Western Blot
All lanes: GIT1 Antibody (Ab105406) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) (Ab170144) at 1/20000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa
Exposure time: 60.0 s

Western Blot: GIT1 Antibody (Ab105406)

GIT1 Antibody (Ab105406) - Western Blot
All lanes: GIT1 Antibody (Ab105406) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) (Ab170144) at 1/20000 dilution
Predicted band size: 84 kDa
Observed band size: 86 kDa
Exposure time: 60.0 s

IHC: GIT1 Antibody (Ab105406)

GIT1 Antibody (Ab105406) - IHC
Immunohistochemical analysis of GIT1 Antibody (Ab105406) staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the GIT1 Antibody (Ab105406) at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Aplicación

Aplicación

Dilution info

WB

1/500-1/1000

IHC

1/50-1/100

Objetivos asociados (humanos)

GIT1 Tbio ARF GTPase-activating protein GIT1 (0 Activities)

Discover Target: GIT1

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

Application Protocols

Manufacturer-tested applications, recommended dilutions, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.

General/literature protocol outlines (for guidance only):

• Western Blot (WB):
  • Prepare lysates with RIPA or mild lysis buffer containing protease/phosphatase inhibitors. Denature with SDS sample buffer; reduce/boil as appropriate.
  • Resolve by SDS-PAGE (10–12%), transfer to PVDF or nitrocellulose.
  • Block 30–60 min, incubate with primary antibody (pilot 0.1–1 µg/mL) 1–2 h RT or overnight 4°C, then with species-appropriate HRP/fluor secondary.
• Immunofluorescence (IF/ICC):
  • Fix cells (e.g., PFA 4%); permeabilize with 0.1–0.3% Triton X-100 if epitope is intracellular.
  • Block and incubate primary (pilot 1–5 µg/mL) 1 h RT or overnight 4°C; wash and add fluorescent secondary.
• Immunoprecipitation (IP):
  • Lyse under gentle conditions for complex preservation. Incubate lysate with antibody (1–5 µg per 0.5–1 mg total protein) and protein A/G beads; wash and elute for WB/MS.

Note: Optimize antibody amounts, incubation times, and buffers empirically. Ensure secondary antibodies match the host/isotype of the primary—host/isotype are not specified for this item.

Biological Roles

Focus: GIT1 (G protein–coupled receptor kinase-interacting protein 1) biology. The antibody is a tool to study these roles; no therapeutic/clinical claims are made.

General/literature overview of GIT1:

• Function: GIT1 is an ARF GTPase-activating protein (ARF-GAP) and scaffold that integrates signaling at focal adhesions and actin cytoskeleton remodeling. It interacts with GRKs, paxillin, PIX/PAK complexes, and contributes to cell migration dynamics.
• Localization: Cytoplasmic with enrichment at focal adhesions and membrane ruffles; capable of shuttling and forming multi-protein complexes influencing endocytosis and receptor trafficking (literature).
• Domains: Contains ARF-GAP domain, ankyrin repeats, Spa2-homology domain (SHD), and paxillin-binding sites, enabling assembly of signaling hubs (literature).
• Modulation: Subject to phosphorylation and other post-translational modifications that regulate binding partners and activity (literature).

Utility of a GIT1 antibody (general):

• Enables detection, quantification, and localization of GIT1 in cells/tissues.
• Facilitates co-immunoprecipitation to map GIT1-centered interactomes.
• Supports studies on adhesion turnover, GPCR desensitization pathways, and cytoskeletal coordination.

Item-specific biological/target details (species reactivity, epitope): Not specified for this item; consult CoA/Spec Sheet.

Buffer Applications

Applicability: Antibodies are typically used within biological buffer systems rather than acting as buffers themselves.

General/literature guidance for buffer systems with antibodies:

• Common working buffers: PBS (10–50 mM phosphate, 150 mM NaCl, pH 7.2–7.4) or TBS (20–50 mM Tris, 150 mM NaCl, pH 7.4–8.0). Choose TBS when phosphate may interfere with downstream assays.
• Blocking and diluents: Use protein-based blockers (e.g., serum, casein) only if compatible with being carrier free and with downstream conjugations; avoid amine-containing buffers for NHS-ester labeling workflows.
• Detergents: Low concentrations of nonionic detergents (e.g., 0.05% Tween-20) can reduce nonspecific binding in immunoassays; verify compatibility with assay readout.
• pH/ionic strength: Maintain near-neutral pH and isotonic salt to preserve antibody conformation and reduce aggregation.

Item-specific buffer composition provided by the manufacturer: Not specified for this item; refer to CoA/Spec Sheet for exact formulation and any preservatives.

Green Alternatives

Applicability: Green solvent alternatives and process-intensification comparisons are not directly applicable to a protein antibody reagent.

Sustainability considerations for antibody use (general guidance):

• Storage efficiency: Aliquot to minimize freeze–thaw cycles and reduce wastage.
• Buffer choices: Use phosphate or TRIS buffers prepared from high-purity salts; avoid hazardous preservatives where possible, consistent with experimental needs and sterility.
• Packaging/cold-chain: Coordinate deliveries to limit dry-ice/ice-pack shipments and consolidate orders when feasible.

There are no greener “chemical substitutes” for a target-specific antibody; however, recombinant production formats (not specified here) can, in general, offer improved batch consistency and potentially reduced animal use.

Pharmaceutical Uses

This product is designated for research use only and is not intended for human or veterinary use, diagnostics, or therapeutic applications.

General/literature context (non-clinical):

• Antibodies like this are commonly used as analytical reagents in bioprocess development, target validation, and mechanism-of-action studies in preclinical research.
• No pharmacopeial monograph or excipient designation is implied for this item. There are no clinical or formulation claims associated with this listing.

Item-specific regulatory/compendial status: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific specifications: Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties for antibodies (context only):

• Approximate molecular mass: ~150 kDa for intact IgG; smaller for fragments (Fab ~50 kDa, scFv ~25–30 kDa) (literature). The exact mass and format for this product are not specified.
• Physical state: Typically supplied as an aqueous protein solution or lyophilized solid. For this item, appearance is not specified.
• pI: Antibodies typically have isoelectric points ranging ~6–9 depending on isotype and glycosylation (literature). Not item-specific.
• Solubility: Antibodies are water-soluble in physiological buffers (e.g., PBS); salt and pH can affect colloidal stability (literature). Not item-specific.
• Stability considerations (literature): Proteins are sensitive to repeated freeze–thaw; stabilizers (e.g., sugars, proteins) are sometimes added. This item is carrier free per Product Data, but no further excipient list is provided.

Refractive index, density, pKa, logP: Not applicable/typically not defined for protein biologics; not specified for this item.

Always consult the product’s CoA/Spec Sheet for exact concentration, format (liquid vs. lyophilized), and any excipients.

Quality and Grades

Item-specific grade notes (from Product Data):

• Carrier Free: Supplied without added carrier proteins (e.g., BSA). This facilitates accurate protein quantitation, conjugation chemistry (e.g., labeling with enzymes/fluorophores), and reduces background in sensitive assays.
• ExactAb™: Provided as an ExactAb™ format. While the vendor branding denotes a high-consistency antibody preparation, clone/isotype and recombinant status are not specified for this item; refer to CoA/Spec Sheet for definitive details.
• Validated: The antibody has undergone validation by the manufacturer; however, the specific tested applications, species reactivity, and validation methods are not listed here.

What this means in practice (general guidance):

• Absence of carrier proteins improves compatibility with mass spectrometry, functional assays, and coupling to solid supports.
• Validation typically includes titer/affinity checks and performance in stated applications. Since this listing does not enumerate them, end users should confirm suitability with small pilot experiments.
• Lot-to-lot consistency and identity are best confirmed using the CoA (e.g., concentration, purity by SDS-PAGE/HPLC, endotoxin if applicable). These metrics are not specified for this item and should be verified on delivery.

Documentation: CoA/Spec Sheet and SDS should be consulted for concentration, buffer composition, and any endotoxin or host/species details.

Reaction and Applications

Chemical reaction applications (synthetic chemistry) are not applicable to antibodies.

Research applications (general/literature guidance for GIT1 antibodies):

• Immunoblotting (WB): Detection of GIT1 protein in cell or tissue lysates to assess expression levels and molecular weight shifts (e.g., post-translational modifications). Optimize antigen retrieval/lysis conditions to preserve GIT1.
• Immunoprecipitation (IP/Co-IP): Enrichment of GIT1 and associated protein complexes (e.g., focal adhesion components) for downstream MS or blot analysis. Use gentle lysis buffers if preserving protein–protein interactions is critical.
• Immunofluorescence/Immunocytochemistry (IF/ICC): Localization of GIT1 at focal adhesions, membrane ruffles, or cytosolic pools in adherent cells; fixation/permeabilization conditions can affect epitope accessibility.
• ELISA/antigen capture: If compatible with the antibody’s epitope, can be used in sandwich ELISAs. Ensure non-overlapping epitopes for capture vs. detection when using paired antibodies.

Item-specific tested applications and species reactivity: Not specified for this item; refer to CoA/Spec Sheet. Users should perform pilot titrations to establish optimal dilutions and blocking conditions.

Reaction Conditions

Not applicable to chemical reactions. This antibody is used under biological assay conditions.

General/literature assay condition guidance for antibodies:

• Working concentration ranges: WB (typically 0.05–1 µg/mL), IF/ICC (0.5–10 µg/mL), IP (1–5 µg per IP), ELISA coating (0.5–2 µg/mL). These are broad literature ranges; optimal conditions must be empirically determined for each lot and assay.
• Temperature/incubation: Many immunoassays perform primary antibody incubations at room temperature for 1–2 h or 4°C overnight to enhance specificity.
• Buffers: PBS or TBS with appropriate blockers and detergents to minimize nonspecific binding; avoid incompatible additives when performing conjugations.

Item-specific tested conditions and recommended dilutions: Not specified for this item; refer to CoA/Spec Sheet. Start with small pilot titrations to optimize signal-to-noise for your specific sample and detection system.

Safety and Handling

Item-specific hazard information:

• GHS Classification, Signal Word, H-Statements, Pictograms: Not specified for this item; refer to the SDS.

General safety guidance for protein antibodies (literature/good practice):

• Expected hazard: Antibodies are generally considered low acute hazard laboratory reagents; nonetheless, handle as potentially irritating biological materials. Avoid inhalation/aerosol generation and contact with skin/eyes.
• PPE: Lab coat, safety glasses, and appropriate gloves (e.g., nitrile). Use biological safety cabinet when preparing sterile aliquots to minimize contamination.
• First aid (overview): If skin/eye contact occurs, rinse with water for several minutes. If inhaled (aerosols), move to fresh air. If ingested, rinse mouth and seek medical advice. Always follow institutional biosafety procedures.
• Incompatibilities/conditions to avoid: Avoid microbial contamination, extreme pH, elevated temperature, and repeated freeze–thaw cycles (can denature protein). Avoid vigorous vortexing/foaming that may cause aggregation.
• Spills/disposal: Absorb small spills with inert material, decontaminate surfaces with suitable disinfectant, and dispose in accordance with local regulations for laboratory biological materials.

Definitive safety information: Defer to the product SDS and institutional EHS guidance. Do not scale beyond research/lab use (Product Data: For research use only).

Solvent Selection

Applicability: As a protein biologic, this antibody is not used as an organic solvent. Solvent selection in the classical sense does not apply.

Relevant buffer considerations (general/literature guidance):

• Typical storage/working buffers: Phosphate-buffered saline (PBS) or other neutral, isotonic buffers at pH ~7.2–7.4 are commonly used for antibodies. For certain applications, Tris-buffered saline (TBS) may be preferred to avoid phosphate interactions.
• Additives: When carrier-free material must remain additive-free for conjugations, avoid introducing proteins (e.g., BSA) or amine-containing buffers if planning NHS-ester labeling. For enzymatic or click conjugations, ensure buffer compatibility (e.g., avoid azide for HRP, avoid primary amines for NHS chemistry).
• Organic co-solvents: Minimize organic solvent exposure; low levels of glycerol are sometimes used to enhance freeze–thaw stability, but this item is carrier free and no excipient list is provided. Confirm buffer composition on the CoA before adding stabilizers.

Item-specific buffer composition and pH: Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific storage and shipping (from Product Data):

• Storage Conditions: Store at -20°C. Avoid repeated freezing and thawing.
• Shipped In: Ice chest + Ice pads.

Additional handling guidance (general best practice; not item-specific):

• Upon receipt: If liquid, briefly centrifuge to collect contents. Inspect for particulates; mix gently—do not vortex vigorously.
• Aliquoting: Prepare small, single-use aliquots to minimize freeze–thaw. Use sterile, low-protein-binding tubes.
• Short-term storage: For frequent use over days to weeks, many antibodies are stable at 2–8°C or -20°C depending on formulation; follow the Product Data (-20°C) and confirm on CoA/Spec Sheet.
• Reconstitution (if lyophilized): Not specified for this item; refer to CoA/Spec Sheet. General practice is to reconstitute with sterile, neutral pH buffer (e.g., PBS) to the desired concentration, then allow to fully dissolve by gentle inversion. Avoid foaming.
• Additives: Since this product is Carrier Free, avoid adding carriers unless required by your application; adding proteins or glycerol may impact downstream conjugations.
• Freeze–thaw guidance: Avoid repeated cycles; keep aliquots on ice during use and return promptly to -20°C.

Definitive instructions (buffer composition, concentration, preservative presence): Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Brief description: GIT1 Antibody is a research-grade immunoreagent directed against the GIT1 protein (G protein–coupled receptor kinase-interacting protein 1), supplied as a carrier-free, validated ExactAb™ format.

Item-specific facts (from Product Data):

• SKU: Ab105406
• Product Name: GIT1 Antibody
• Grade/Purity: Carrier Free; ExactAb™; Validated
• CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Formula, Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.

General/literature notes (antibody identity):

• Type/Structure (literature): Typical IgG-class antibodies are Y-shaped glycoproteins composed of two heavy and two light chains with disulfide linkages; intact IgG has an approximate mass of ~150 kDa (literature), with variable domains (Fab) mediating antigen binding and an Fc domain mediating effector functions. Exact subclass/isotype for this item is not specified.
• Antigen: GIT1 (also known as ARF GTPase-activating protein GIT1), a scaffold and ARF-GAP protein involved in focal adhesion dynamics and signaling. Epitope/peptide region not specified for this item.

2D/structural features (descriptive, literature): Antibodies present conserved Ig domains (β-sandwich folds) in both heavy and light chains. The paratope resides in hypervariable complementarity-determining regions (CDRs). No item-specific structural characterization is provided.

Synthetic Utility

Not applicable. As a protein biologic, the GIT1 Antibody is not used as a synthetic building block or reagent for organic transformations.

General/literature utility in analytical/biochemical workflows:

• Target recognition tool enabling immunoassays (WB, ELISA), immunocapture for proteomics, and imaging (IF/ICC/IHC) studies.
• Can be conjugated to enzymes, fluorophores, biotin, or solid supports to create affinity reagents, provided buffer components are compatible with the chosen conjugation chemistry.

Item-specific conjugation readiness: The product is Carrier Free, which is advantageous for chemical conjugation. Specific buffer composition and reactive groups available are not specified for this item; verify on CoA/Spec Sheet before performing labeling.

Target Specificity

Item-specific details (from Product Data):

• Antigen/Target: GIT1 (G protein–coupled receptor kinase-interacting protein 1)
• Clone, isotype, host species, epitope/peptide, species reactivity, cross-reactivity: Not specified for this item; refer to CoA/Spec Sheet.

General/literature context:

• GIT1 is a conserved scaffold/ARF-GAP protein implicated in focal adhesion signaling and cytoskeletal regulation. Antibodies may recognize linear or conformational epitopes depending on design and validation (not specified here).

Recommendation: Confirm species reactivity and epitope region on the CoA/Spec Sheet before committing to large studies. Validate specificity in your system with positive and negative controls (e.g., siRNA/CRISPR knockdown/knockout of GIT1, or overexpression).

Preguntas frecuentes

What does ExactAb? mean?

ExactAb? indicates the product has been validated for use in rigorous antibody validation workflows. Validation may include positive/negative control samples, signal-to-noise testing, and protocol-specific QC. Lot-specific test data is published on the Certificate of Analysis where available.

What does Carrier Free mean?

Carrier Free indicates that the material has been processed and tested for specialty industrial processing. Specifications focus on the impurities that affect performance in that application — for example, metal content for electronics, particle size for batteries, refractive index for optics, or moisture content for moisture-sensitive processes.

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.

Which species reactivity has this antibody been validated against?

Validated reactivity is listed on the PDP and on each lot's datasheet. We validate against the most commonly tested species for the target (GIT1 Antibody); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.