ZAP70 mAb di topo - Anticorpo primario, specific to ZAP70, Mouse IgG

Cat. No.: Ab135097

QUALITÀ & PUREZZA Carrier Free, Azide Free, Validated, 1.0 mg/mL

Application

IHC

Host species

Mouse

Reactive species

Umano

Isotype

Mouse IgG

Clone Number

YN01716m

Conjugation

Unconjugated

Panoramica

L'anticorpo di topo anti-ZAP70 umano, monoclonale (YN01716m), può essere utilizzato per IHC e così via.
Applicazione
IHC: 1/200
Funzione della proteina
Svolge un ruolo nello sviluppo delle cellule T e nell'attivazione dei linfociti. È essenziale per la produzione di IL-2 mediata dal TCR. L'isoforma 1 induce la trasduzione del segnale mediata dal TCR, mentre l'isoforma 2 non lo fa.

Specifiche

Product Name

ZAP70 mAb di topo - Anticorpo primario, specific to ZAP70, Mouse IgG

Sinonimi

70 kDa zeta associated protein anticorpo | 70 kDa zeta-associated protein anticorpo | EC 2.7.10.2 anticorpo | FLJ17670 anticorpo | FLJ17679 anticorpo | Selective T cell defect anticorpo | SRK anticorpo | STD anticorpo | Syk related tyrosine kinase anticor

Specifiche e purezza

Privo di azide, convalidato, privo di carrier, 1,0 mg/mL

Specie ospite

Mouse

Specificità

ZAP70

Immunogeno

Peptide

Controllo positivo

IHC: Human tonsil tissue.

Coniugazione

Unconjugated

Grado

Azide Free, Carrier Free, Validated

Isotipo

Mouse IgG

Tipo di catena leggera

kappa

SDS-PAGE

150 kDa

Metodo di purificazione

Protein G purified

Concentrazione

1.0 mg/mL

Product Line

Prodotti biologici

Stoccaggio e spedizione

Forma

Liquido

Condizioni di conservazione di stoccaggio

Conservare a -20°C, evitando di congelare e scongelare ripetutamente

Spedito in

Cassetta del ghiaccio + cuscini per il ghiaccio

Stabilità e conservazione

Conservare a -20℃ a lungo termine (12 mesi). Al momento del ricevimento, si consiglia di effettuare un'aliquota. Evitare il ciclo di congelamento/scongelamento.

Immagini

IHC: ZAP70 Mouse mAb (Ab135097)

ZAP70 Mouse mAb (Ab135097) - IHC
Immunohistochemistry analysis of paraffin-embedded human tonsil tissue labelling ZAP70 with ZAP70 Mouse mAb (Ab135097) at 1/200 dilution. The section was incubated with ZAP70 Mouse mAb (Ab135097) for 30 mins at room temperature. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was used for antigen retrieval.​

IHC: ZAP70 Mouse mAb (Ab135097)

ZAP70 Mouse mAb (Ab135097) - IHC​
Immunohistochemistry analysis of paraffin-embedded human tonsil tissue labelling ZAP70 with ZAP70 Mouse mAb (Ab135097) at 1/200 dilution. The section was incubated with ZAP70 Mouse mAb (Ab135097) for 30 mins at room temperature. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was used for antigen retrieval.​

Applicazione

Applicazione

Dilution info

IHC

1/200

Obiettivi associati (umani)

ZAP70 Tchem Tyrosine-protein kinase ZAP-70 (0 Activities)

Discover Target: ZAP70

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

Meccanismi d'azione

Application Protocols

Item-specific, tested/validated applications and recommended working conditions are Not specified for this item; refer to CoA/Spec Sheet.

General protocol outlines (for planning only; not item-specific)

• Western blot: Resolve lysates from T‑cell lines (e.g., Jurkat) or stimulated primary T cells; transfer to PVDF; block; incubate with anti‑ZAP70 primary; wash; detect with species‑appropriate secondary.
• Immunoprecipitation: Lyse cells under mild, non‑denaturing conditions; incubate lysate with the antibody, capture with protein A/G resin (isotype-dependent); wash; elute and analyze by WB.
• Immunofluorescence/ICC: Fix and permeabilize cells (PFA + Triton or methanol as appropriate); block; incubate with primary; detect with fluorescent secondary; image.
• Flow cytometry: For intracellular staining, fix/permeabilize T cells; stain with primary and labeled secondary; include isotype and FMO controls.

Critical reminders

• Because clone, isotype, concentration, and dilution ranges for SKU Ab135097 are not provided here, optimize empirically and consult the CoA/Spec Sheet before use.

Biological Roles

General/literature context for the target ZAP70 (not item-specific)

• Function: ZAP70 (Zeta-chain-associated protein tyrosine kinase 70) is a cytoplasmic tyrosine kinase essential for T‑cell receptor (TCR) signaling. Upon TCR engagement, ZAP70 is recruited to phosphorylated ITAMs on CD3ζ via its tandem SH2 domains, becomes activated (via phosphorylation by Lck and autophosphorylation), and phosphorylates downstream scaffolds (LAT, SLP‑76), propagating calcium influx, MAPK activation, and transcriptional programs.
• Expression: Predominantly in T cells and natural killer (NK) cells; also detectable in certain B‑cell malignancies where it has prognostic relevance (literature).
• Cellular localization: Cytoplasmic; transiently associates with the plasma membrane at the immune synapse following TCR activation.
• Pathways: TCR signaling, NK cell–mediated cytotoxicity, immune synapse assembly, and development of adaptive immunity.
• Structural notes: Contains tandem SH2 domains for phosphotyrosine recognition and a C‑terminal kinase domain (TK). Key regulatory tyrosines include Y319 (adaptor function) and activation loop residues (Y493, Y492) (literature).
• Research utility: Antibodies to ZAP70 are used for detection of total protein levels, assessment of activation state (using phospho‑specific clones), and immunoprecipitation of signaling complexes.

Item-specific details such as epitope, species cross‑reactivity, and application validations are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

Applicability to antibodies

• This product is a protein reagent rather than a buffering agent; it does not establish pH. However, successful use depends on choosing suitable buffers for the intended immunoassay.

General buffer guidance (not item-specific)

• Western blot/IHC/IF: PBS or TBS at pH 7.2–7.6 with 0.05% Tween‑20 for washes; blocking with 3–5% BSA or milk (if compatible). For phospho‑epitope work, prefer BSA to avoid phosphoprotein interference.
• Flow cytometry: PBS + 1–2% BSA + 0.05% sodium azide is common; for live cells, omit azide (this product is azide‑free as supplied).
• Immunoprecipitation: Isotonic buffers (e.g., 20–50 mM Tris, 150 mM NaCl, 0.1% NP‑40 or Triton X‑100) with protease/phosphatase inhibitors tailored to preserve ZAP70 complexes.
• Antigen retrieval (IHC): Citrate buffer pH 6.0 or Tris‑EDTA pH 9.0 can unmask epitopes; optimize per tissue and fixation.

Item-specific buffer formulation and recommended working dilutions: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Conventional “green alternatives” guidance typically compares organic solvents or reagents; for a monoclonal antibody, the sustainability focus centers on formulation, use, and waste minimization rather than chemical substitution.

Greener practice options (general)

• Buffer selection: Use phosphate or HEPES buffers prepared from concentrated stocks to minimize packaging waste. Avoid azide where downstream biological systems are impacted—this item is already Azide Free.
• Stabilization to reduce waste: Aliquot to prevent freeze–thaw loss; employ low‑bind consumables to reduce adsorption losses and re‑runs.
• Reusability: Couple the mAb to solid supports (e.g., NHS‑activated resins) for repeated immunoprecipitation cycles, reducing per‑assay antibody consumption.
• Energy: Store at the lowest recommended temperature (−20°C for this product) rather than ultra‑low where not required to reduce energy footprint, while maintaining stability.
• Packaging: Consolidate orders and select appropriate pack sizes to match consumption and minimize cold‑chain shipments.

Trade‑offs

• Preservatives (e.g., azide, ProClin) can extend shelf life and reduce wastage but may introduce environmental or assay compatibility concerns. This product’s azide‑free status favors biosafety and waste simplification but necessitates stricter aseptic technique.

Pharmaceutical Uses

Scope limitation

• This product is supplied strictly for research use only (per Product Data) and is not intended for diagnostic, therapeutic, or clinical use.

Formulation/excipient context (general; not item-specific)

• Monoclonal antibodies in pharmaceutical manufacturing are therapeutic actives, not excipients. For laboratory settings, antibodies may be used as process development reagents, reference materials, or analytical controls.
• Azide-free, carrier-free status can be advantageous where downstream bioprocess assays or conjugations require minimal additives and low background.

Standards and compliance

• Pharmacopeial monographs do not generally apply to research antibodies of undefined clone/isotype. Any GMP suitability, sterility validation, bioburden, or endotoxin specifications are Not specified for this item; refer to CoA/Spec Sheet.

Regulatory note

• Do not use in humans or animals for therapeutic or diagnostic purposes. Institutional biosafety approval may be required for certain applications.

Physical Properties

Item-specific properties

• Appearance, formulation, concentration, buffer, pH: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight (exact), extinction coefficient, isotype: Not specified for this item; refer to CoA/Spec Sheet.

General/literature values for mouse IgG (not item-specific)

• Approximate molecular weight: ~150 kDa for intact IgG; ~50 kDa heavy chain and ~25 kDa light chain under reducing SDS–PAGE.
• Isoelectric point (pI): Commonly in the 6–9 range depending on isotype and glycosylation.
• Solubility: High in aqueous buffers (e.g., PBS, TBS); insoluble in most organic solvents.
• Stability modifiers: Addition of protein carriers (e.g., BSA), sugars (e.g., trehalose), and mild detergents (e.g., 0.01% Tween‑20) can improve stability; however, this item is designated Carrier Free—do not add carriers unless required by your application.
• Temperature sensitivity: Repeated freeze–thaw cycles can cause aggregation and loss of activity (literature).

Spectral/optical

• UV absorbance: Proteins typically A280 with ε ≈ 1.35–1.6 (mg/mL)−1 cm−1 for IgG (literature). Exact value for this mAb is not specified for this item; refer to CoA/Spec Sheet.

Handling implication

• As an azide‑free product, there is no preservative contribution to ionic strength or UV background from NaN3.

Quality and Grades

Item-specific grades (from Product Data)

• Carrier Free: Supplied without protein carriers (e.g., BSA, gelatin). Advantages include lower background in immunoassays, cleaner conjugation chemistry, and defined mass balance. Caution: Greater sensitivity to surface adsorption—pre-block plastics or add a compatible carrier to working solutions if required by your protocol.
• Azide Free: No sodium azide preservative—compatible with live-cell applications and peroxidase-based detection systems. Caution: Increased microbial risk; maintain sterility and consider single-use aliquots.
• Validated: The product has undergone internal validation by the manufacturer. Specific validated applications, species reactivity, and performance metrics are Not specified for this item; refer to CoA/Spec Sheet.

What this means in practice

• Analytical expectations: For carrier-free mAbs, low non-protein background and predictable conjugation stoichiometry (e.g., for HRP, fluorophores, biotin) are typical benefits (general).
• Documentation check: For clone, isotype, concentration, purity method (e.g., SEC-HPLC, SDS–PAGE), endotoxin level, and residual host cell proteins/DNA, consult the CoA/Spec Sheet—Not specified for this item.

Comparison to other grades (general)

• Carrier-containing formats may increase stability during storage/handling but can confound mass-based dosing and downstream conjugations; azide-containing buffers extend shelf life but are incompatible with certain assays and cell systems.

Reaction and Applications

This section primarily addresses small‑molecule chemical reactivity and is not directly applicable to monoclonal antibodies.

Relevant applications for this product type (general; not item-specific)

• Immunodetection: Western blot (WB), immunohistochemistry (IHC), immunofluorescence (IF), immunocytochemistry (ICC), flow cytometry (FC), and immunoprecipitation (IP) targeting ZAP70.
• Conjugations: Site‑nonspecific labeling via primary amines (NHS esters) or sulfhydryls (maleimides) for HRP, AP, fluorophores, or biotin. For carrier‑free antibodies, conjugation stoichiometry is easier to control.
• Capture/detection: Sandwich ELISA as capture or detection antibody when paired with a non‑overlapping anti‑ZAP70 clone (literature best practice).

Item-specific, manufacturer‑validated applications: Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Classical reaction conditions (solvent, temperature, catalysts) are not applicable to monoclonal antibodies as they are not chemical reagents for synthetic transformations.

Assay/handling conditions (general; not item-specific)

• Incubation: Most immunoassays use 30–120 min at room temperature or overnight at 2–8°C for primary antibody binding; optimize per assay.
• Temperature: Maintain on ice or at 2–8°C during cell staining or IP setup to preserve complexes; avoid elevated temperatures unless protocol‑specific.
• Concentration/dilutions: Typical working dilutions vary widely (e.g., 1:500–1:5000 for WB, 0.5–5 µg/mL for IF/FC), but exact values for this item are Not specified for this item; refer to CoA/Spec Sheet.
• Detergents/salts: Choose based on epitope exposure and background; excessive detergent or salt may disrupt protein–protein interactions relevant to ZAP70 complexes.

Validation

• Manufacturer‑validated conditions, species, and positive control materials are Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

GHS/SDS

• GHS classification, pictograms, hazard and precautionary statements: Not specified for this item; refer to the product SDS for authoritative safety information.

General safety guidance for protein antibodies (literature/practice)

• Expected hazards: Antibodies in research formulations are generally considered low hazard; nonetheless, treat as laboratory reagents of unknown toxicity.
• Personal protective equipment (PPE): Lab coat, nitrile gloves, and safety glasses. Avoid skin/eye contact and inhalation of aerosols.
• Engineering controls: Work in a clean area; for procedures generating aerosols (e.g., sonication, spraying), use a biosafety cabinet if appropriate to your facility’s biosafety assessment.
• Hygiene: Do not pipette by mouth. Wash hands after handling. Decontaminate surfaces with appropriate protein-compatible cleaners.

Incompatibilities and special considerations

• Preservatives: This item is Azide Free—beneficial for peroxidase-based assays and for cells sensitive to azide; however, lack of preservative can increase risk of microbial growth. Maintain sterility and consider sterile filtration of working solutions.
• Metals and plumbing: Azide-free status eliminates the risk of metal azide formation in drains.
• Freeze–thaw: Avoid repeated freeze–thaw cycles to minimize aggregation; aliquot upon first use.

First aid (general)

• Skin/eye contact: Rinse with water. Seek medical attention if irritation persists.
• Ingestion/inhalation: Rinse mouth, move to fresh air; obtain medical advice.

Waste

• Dispose of proteinaceous laboratory waste according to institutional and local regulations; do not discharge to sink unless permitted.

Solvent Selection

Applicability

• As a protein biologic, this monoclonal antibody is intended for aqueous buffers, not organic solvents.

Buffer considerations (general guidance; not item-specific)

• Compatible systems: Phosphate-buffered saline (PBS), Tris-buffered saline (TBS), HEPES buffers at physiological ionic strength (100–200 mM total salts), pH 6.8–8.0 depending on application.
• Stabilizers: For working solutions, addition of 0.05–0.1% carrier protein (e.g., BSA) and/or 2–5% sugar (e.g., trehalose, sucrose) can reduce adsorption and freeze–thaw stress. Note: This product is Carrier Free as supplied; add stabilizers only if appropriate for your assay.
• Detergents: 0.01–0.05% nonionic detergent (Tween‑20) can reduce nonspecific binding in immunoassays.
• Preservatives: If long-term storage at 2–8°C is necessary, non-azide preservatives (e.g., ProClin, sodium benzoate) may be considered where compatible; verify impact on your assay.

When to choose PBS vs TBS

• PBS: Common for Western blot, flow cytometry, and general storage; interacts minimally with most proteins but may interfere with some phospho-specific interactions.
• TBS: Preferred in certain immunoblots to reduce background with phosphate-sensitive interactions or alkaline phosphatase detection.

Incompatibilities

• Avoid high concentrations of organic solvents, strong acids/bases, and chaotropes unless intentionally used for elution/regeneration; these can denature or aggregate the antibody.

Item-specific formulation (buffer, pH, excipients): Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific instructions (from Product Data)

• Storage: Store at −20°C. Avoid repeated freezing and thawing.
• Shipping: Shipped in an ice chest with ice pads (cold‑chain maintained).

Additional guidance (general; not item-specific)

• Upon receipt: If supplied in liquid form, mix gently by inversion, briefly centrifuge to collect contents, and aliquot into low‑bind tubes to avoid multiple freeze–thaw cycles. Freeze aliquots at −20°C or below per datasheet guidance.
• If lyophilized: Reconstitute with sterile, antibody‑grade water or buffer (e.g., PBS) to the desired concentration as recommended in the CoA/Spec Sheet; allow to dissolve gently without vigorous vortexing. After reconstitution, aliquot and store at −20°C.
• Working solutions: Prepare in sterile PBS or TBS with optional stabilizers (e.g., 0.1% BSA, 2–5% trehalose) if compatible with your assay. Discard contaminated or turbid solutions.
• Freeze–thaw management: Thaw on ice, keep cold during use, and refreeze promptly. Avoid repeated cycles to minimize aggregation and activity loss.

Not specified for this item; refer to CoA/Spec Sheet

• Exact formulation/buffer composition and pH
• Protein concentration and volume
• Preservatives/excipients (other than azide‑free designation)
• Long‑term stability claims or expiry date

Structure and Identity

Item-specific facts (from Product Data)

• Product name: ZAP70 Mouse mAb (SKU: Ab135097)
• Type: Monoclonal antibody, host species: Mouse
• Grade/Purity: Carrier Free, Azide Free, Validated
• Target: ZAP70 (Zeta-chain-associated protein kinase 70)
• CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.

General/literature description (antibody architecture; not item-specific)

• Typical antibody class: IgG monoclonal antibodies are Y‑shaped glycoproteins composed of two identical heavy chains and two identical light chains, each with variable (V) and constant (C) domains.
• Structural features: Two antigen-binding fragments (Fabs) with complementary determining regions (CDRs) recognizing an epitope on ZAP70; one Fc region mediating effector and stability properties. Disulfide bonds link heavy–heavy and heavy–light chains; N‑glycosylation commonly present in Fc.
• Molecular formula/weight: Not defined for a heterogeneous glycoprotein; typical intact IgG MW ~145–155 kDa (literature).
• 2D description in words: A symmetric Y‑shaped heterotetramer with paired β‑sheet immunoglobulin folds forming variable domains at the tips (paratopes) and constant domains forming the stem (Fc). Epitope specificity is determined by the CDR loops of the VH/VL domains.

Notes on identity control

• Clone identifier, isotype, immunogen, and epitope mapping: Not specified for this item; refer to CoA/Spec Sheet.
• Sequence information (VH/VL or recombinant format): Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not applicable to this product type.

Explanation

• “Synthetic utility” refers to the role of small molecules as building blocks, reagents, or catalysts in chemical synthesis. Monoclonal antibodies are biological macromolecules used for detection, capture, or modulation of targets in biochemical assays rather than as reagents in organic synthesis.

Adjacent relevant capabilities (general)

• Antibody–drug conjugate (ADC) research: Carrier‑free antibodies can be chemically modified (e.g., NHS‑ester, maleimide, click chemistry) to generate conjugates for in vitro studies.
• Affinity matrices: Coupling to activated resins (e.g., NHS Sepharose) to construct immunoaffinity columns for purification or target enrichment.

Target Specificity

Item-specific (from Product Data)

• Target antigen: ZAP70 (Zeta-chain-associated protein kinase 70)
• Antibody type/host: Mouse monoclonal antibody (mAb)

Not specified for this item; refer to CoA/Spec Sheet

• Clone ID and isotype
• Epitope/immunogen details (e.g., peptide vs recombinant protein region)
• Species reactivity and cross‑reactivity profile
• Affinity/avidity (KD), dissociation rates
• Validated applications and positive control recommendations
• Reactivity with post‑translationally modified forms (e.g., phospho‑specificity)

General note

• ZAP70 resides primarily in T cells/NK cells and associates with TCR signaling complexes; verify species and tissue reactivity in your system with appropriate controls.