UBE2E3 Anticorpo - Anticorpo primario, specific to UBE2E3, Rabbit IgG

Cat. No.: Ab133237

QUALITÀ & PUREZZA See COA

Application

Flow Cytometry, IF/ICC, IHC, WB

Host species

Mouse

Reactive species

consiglio, Mouse, Scimmia, Tuttavia, Umano

Isotype

Mouse IgG1

Clone Number

OTI1D12

Conjugation

Unconjugated

Panoramica

L'anticorpo di coniglio anti UBE2E3 umano, policlonale, può essere utilizzato per WB, ICC, IF e così via.
Applicazione:
WB: 1/500-1/1000

IF/ICC: 1/50-1/200

Funzione della proteina

Accetta l'ubiquitina dal complesso E1 e ne catalizza il legame covalente con altre proteine. In vitro catalizza la poliubiquitinazione legata a 'Lys-11' e 'Lys-48', nonché a 'Lys-63'. Partecipa alla regolazione del trasporto transepiteliale del sodio nelle cellule renali. Può essere coinvolto nell'arresto della crescita cellulare.

Specifiche

Product Name

UBE2E3 Anticorpo - Anticorpo primario, specific to UBE2E3, Rabbit IgG

Sinonimi

UbcM2 | UBCH9 | UBE2E3 | Ubiquitin-protein ligase E3 antibody | Ubiquitin-conjugating enzyme E2-23 kDa antibody | UB2E3_HUMAN antibody | Homologous to yeast UBC4/5 antibody | UBC E4 antibody | Ubc M2 antibody | UBC4/5 homolog antibody | Ubiquitin-conjugat

Specifiche e purezza

ExactAb™, convalidato, senza carrier, 1,0 mg/mL

Meccanismi biochimici e fisiologici

Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro catalyzes 'Lys-11'- and 'Lys-48'-, as well as 'Lys-63'-linked polyubiquitination. Participates in the regulation of transepithelial sodium transport in

Specie ospite

Mouse

Specificità

UBE2E3

Immunogeno

Recombinant protein of human UBE2E3 (AA 1-207).

Controllo positivo

WB: Hep-G2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12, MCF7 lysates. IHC: Human colon, kidney, ovary, pancreas, thyroid, endometrium, prostate, bladder tissue. IF/ICC: HEK293E cells. Flow Cytometry: HEK293T cells.

Coniugazione

Unconjugated

Isotipo

Rabbit IgG

SDS-PAGE

150 kDa

Metodo di purificazione

Protein A/G purified

Concentrazione

See COA

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 4℃ a breve termine (1-2 settimane). Conservare a -20℃ a lungo termine (12 mesi). Al momento della consegna, aliquottare. Evitare il ciclo di congelamento/scongelamento.

Applicazione

Applicazione

Dilution info

WB

1/500-1/2000

Flow Cytometry

1/100

IHC

1/150

IF/ICC

1:100

Obiettivi associati (umani)

UBE2E3 Tbio Ubiquitin-conjugating enzyme E2 E3 (0 Activities)

Discover Target: UBE2E3

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

Application Protocols

Manufacturer-validated applications and recommended dilutions for Ab133237 are not specified for this item; refer to CoA/Spec Sheet. The following are general starting points (literature; not product-validated) for mouse monoclonal antibodies:

• Western blot (WB): 1:1,000 to 1:5,000 in TBST with 5% BSA or milk; incubate 1 h RT or overnight 4°C. Load 10–40 µg total protein per lane; use reducing and denaturing conditions unless epitope is reduction-sensitive.
• Immunofluorescence/ICC: 1:100 to 1:500 in blocking buffer; incubate 1 h RT or overnight 4°C. Include permeabilization (e.g., 0.1–0.2% Triton X-100) for intracellular targets.
• Immunohistochemistry (IHC-P): 1:100 to 1:400 after antigen retrieval (citrate pH 6.0 or Tris-EDTA pH 9.0). Block endogenous peroxidase for HRP detection.
• Immunoprecipitation (IP): 1–5 µg antibody per 0.5–1 mg lysate protein; incubate 1–4 h at 4°C; capture with Protein A/G beads appropriate for the isotype.
• ELISA (indirect): 0.1–1 µg/mL primary; optimize coating and blocking conditions to minimize background.

Controls and validation

• Use isotype-matched control IgG and secondary-only controls.
• Confirm band size and specificity with siRNA/CRISPR knockdown where feasible.
• Optimize antigen retrieval, blocking, and washing stringency per sample type.

Biological Roles

Target overview (general, literature)

• UBE2E3 is a member of the ubiquitin-conjugating enzyme (E2) family. E2s receive activated ubiquitin from E1 enzymes and collaborate with E3 ligases to transfer ubiquitin to substrate proteins.
• Functional scope: E2 enzymes influence the topology of ubiquitin chains (mono- vs polyubiquitination; Lys-linkage preferences) in combination with partner E3s, thereby regulating substrate stability, trafficking, and activity.
• Cellular context: UBE2E3 is reported to localize to cytosolic and nuclear compartments depending on cell type and conditions. Its expression is part of the broader ubiquitin–proteasome system (UPS), central to proteostasis.
• Pathway integration: Ubiquitination modulates cell cycle progression, DNA damage responses, signaling cascades, and protein quality control (general roles of the UPS; specific pathways for UBE2E3 vary by system).

Why target UBE2E3 experimentally (general)

• Protein homeostasis studies: Monitoring UBE2E3 abundance/localization can report on UPS competence and stress responses.
• Interaction mapping: IP mass spectrometry can define UBE2E3-associated E3 ligases and substrates in a given model.
• Perturbation analyses: siRNA/CRISPR knockdown/knockout and rescue experiments combined with antibody-based readouts enable functional attribution in ubiquitination pathways.

Note: Species reactivity, epitope, and affinity of Ab133237 are not specified for this item; refer to CoA/Spec Sheet for product-specific details before designing experiments.

Buffer Applications

Direct buffer-formulation roles do not apply to this product in the classical sense (it is not a buffering agent). However, buffer choice is critical for antibody performance.

General guidance for antibody workflows (not item-specific)

• Storage buffers: Neutral pH buffers (PBS or TBS) with potential stabilizers like 10–50% glycerol and carrier protein (e.g., 0.1% BSA) can enhance freeze–thaw resilience; composition for Ab133237 is not specified.
• Assay buffers: For WB/ELISA/IF, use PBS/TBS with 0.05–0.1% Tween-20 for washing to minimize nonspecific binding; block with 3–5% BSA or nonfat milk depending on detection chemistry.
• Antigen retrieval (IHC): Heat-induced epitope retrieval in citrate (pH ~6.0) or Tris-EDTA (pH ~9.0) buffers may be required; optimize per tissue and fixation.

Compatibility warnings

• Avoid amine-containing buffers (e.g., Tris) during NHS-ester labeling chemistry.
• If preservatives like sodium azide are present (not specified here), they can inhibit HRP; use azide-free buffers for HRP-based detection.

For Ab133237, exact buffer formulation and compatibility are not specified; consult the CoA/Spec Sheet.

Green Alternatives

Antibody-related sustainability considerations (general)

• Minimize waste: Order pack sizes aligned with experimental throughput to reduce expired material.
• Storage efficiency: Use aliquots to avoid freeze–thaw losses; this reduces repeat purchases and waste generation.
• Buffer selection: Use phosphate or Tris buffers prepared from concentrated stocks to reduce packaging; avoid hazardous preservatives where feasible and compatible with your biosafety requirements.
• Detection systems: Prefer HRP substrates with safer chemistries or fluorescent detection to reduce solvent-intensive film development.
• Reusable materials: Adopt reusable gel cassettes and staining trays to cut single-use plastics.

Alternative reagent formats (trade-offs, general)

• Recombinant monoclonal antibodies: Offer lot-to-lot consistency and defined sequences; may reduce animal use, improving ethical and sustainability profiles.
• Antibody fragments (Fab/F(ab’)2, scFv): Smaller formats can improve tissue penetration and reduce background in certain assays, but may have shorter half-life and require tailored secondary reagents.

Note: No greener “chemical substitute” applies here, as this is a biologic. Product-specific stabilizers/preservatives for Ab133237 are not specified; consult the CoA/Spec Sheet to assess any hazardous components and choose disposal practices accordingly.

Pharmaceutical Uses

This product is designated for research use only and is not intended for human or veterinary use, diagnostic procedures, or as a therapeutic.

Relevant non-clinical contexts (general)

• Process analytics: Research-grade antibodies can be used in process development assays (e.g., Western blot or ELISA) to monitor target protein expression in cell lines during upstream development.
• Reagent for assay development: As a capture or detection reagent in prototype immunoassays for preclinical research.

Regulatory considerations

• No pharmacopeial status or GMP qualification is indicated for this item. Any use beyond basic research (e.g., QC release testing) requires appropriate qualification/validation.

Item-specific statement

• Grade/purity, endotoxin, sterile filtration status, and preservative content are not specified for this item; refer to CoA/Spec Sheet if considering regulated workflows.

Physical Properties

Item-specific physical data

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer composition, preservatives: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight (exact): Not specified for this item; refer to CoA/Spec Sheet.

General properties of mouse IgG monoclonal antibodies (literature/general)

• Approximate molecular mass: ~150 kDa for intact IgG; Fab fragments ~50 kDa; Fc ~50 kDa; scFv ~25–30 kDa (if applicable; format for this item not specified).
• Solubility: Readily soluble in aqueous physiological buffers (e.g., PBS, TBS). Insoluble in most organic solvents.
• Isoelectric point (pI): Commonly ~6–9 depending on isotype and sequence.
• Extinction coefficient: Sequence-dependent; typical A280 conversion factors for IgG are ~1.3–1.6 (mg/mL)−1 cm−1, but item-specific value is not provided here.
• Stability: Proteins are heat-labile; avoid elevated temperatures and repeated freeze–thaw.
• Melting/boiling point: Not applicable for proteins; denaturation occurs well below water’s boiling point.

Practical notes

• Gentle handling minimizes aggregation; avoid vigorous vortexing.
• If supplied with glycerol (not specified here), viscosity increases at low temperature; equilibrate on ice before pipetting.

Quality and Grades

Item-specific quality information

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Tested applications, species reactivity, clone/isotype, purification method (Protein A/G, antigen affinity), endotoxin/bioburden levels: Not specified for this item; refer to CoA/Spec Sheet.

How to interpret antibody quality (general guidance)

• Monoclonal antibodies (mAbs) provide epitope-specific binding and lot-to-lot consistency compared with polyclonals.
• Typical QC for research antibodies can include: SDS-PAGE (purity/HC-LC pattern), SEC-HPLC (aggregate content), ELISA/SPR (affinity), application-specific validation (WB, IHC, IF, IP), and species cross-reactivity panels.
• Isotype matters for application choice (e.g., IgG1 vs IgG2a for Fc interactions and secondary selection); isotype for this item is not specified.
• Purification methods influence performance: Protein A/G yields high purity; antigen affinity can enrich for high-specificity populations; details not specified for this item.
• Low endotoxin is important for cell culture work; value not specified here.

Stabilizers and preservatives

• Common stabilizers include BSA, trehalose, and glycerol; presence/absence for this item is not specified. Stabilizers can affect downstream assays (e.g., BSA can interfere with protein labeling or ELISA background). Always check the CoA/Spec Sheet for exact formulation.

Reaction and Applications

This section focuses on practical research applications rather than chemical reactions, given the product is an antibody.

Typical research applications for a UBE2E3-targeting mouse mAb (general; not validated for this specific item)

• Western blotting (WB): Detection of endogenous UBE2E3 in cell/tissue lysates to assess expression changes upon pathway perturbation (e.g., proteostasis, ubiquitination studies).
• Immunoprecipitation (IP)/Co-IP: Enrichment of UBE2E3 to study interacting E3 ligases or substrate complexes.
• Immunofluorescence (IF)/Immunocytochemistry (ICC): Subcellular localization of UBE2E3 (cytosolic/nuclear distribution) under different conditions.
• Immunohistochemistry (IHC): Spatial mapping of UBE2E3 in tissues.
• ELISA/Target capture: Development of sandwich or indirect ELISAs if epitope accessibility is suitable.

Experimental considerations (general)

• Epitope sensitivity: Denaturation (WB) vs native (IP/IF) conditions can expose or mask epitopes; optimization of antigen retrieval and reduction/boiling is critical.
• Controls: Include isotype controls, peptide competition (if immunizing peptide is available), and knockout/siRNA controls where feasible.
• Signal optimization: Titrate antibody, optimize blocking (e.g., 3–5% BSA or milk; selection depends on detection chemistry), and choose appropriate secondary antibodies.

Manufacturer-validated applications and dilutions for Ab133237 are not specified for this item; consult the CoA/Spec Sheet for product-specific recommendations.

Reaction Conditions

Classical chemical reaction conditions do not apply. For immunoassay workflows, the following general incubation conditions are commonly used for mouse monoclonal antibodies (literature/general guidance; not validated for Ab133237):

• Western blot primary incubation: 1–16 h at 4–22°C in TBST/PBST with 3–5% BSA or milk. Longer, colder incubations can improve specificity; optimize antibody dilution to balance signal and background.
• Immunofluorescence/ICC: 1 h at room temperature or overnight at 4°C in blocking buffer. Permeabilize with 0.1–0.3% Triton X-100 if targeting intracellular antigens; optimize to preserve epitope.
• Immunohistochemistry: After antigen retrieval, incubate 1 h RT or overnight 4°C; blocking with serum or protein blocker reduces background.
• Immunoprecipitation: 1–4 h at 4°C (end-over-end) in lysis buffer (e.g., NP-40 or Triton-based) with protease inhibitors; capture with Protein A/G resin appropriate for the antibody isotype.

Buffers, detergent types, and retrieval protocols must be tailored to the sample and detection system. Product-specific optimal dilutions and conditions for Ab133237 are not specified; consult the CoA/Spec Sheet and perform preliminary titration/optimization.

Safety and Handling

GHS/SDS

• GHS Classification, Signal Word, H-Statements, Pictograms: Not specified for this item; consult the product SDS for authoritative hazard information.

General laboratory safety for antibodies (advisory)

• PPE: Laboratory coat, gloves (nitrile/latex), and safety glasses as standard practice.
• Biosafety: Treat as a biological reagent; avoid ingestion and contact with skin/eyes. Handle using aseptic technique if sterility is required.
• Preservatives: Presence/absence of sodium azide, ProClin, or other preservatives is not specified for this item. If azide is present, avoid contact with metals (e.g., copper/lead) to prevent formation of metal azides; flush drains thoroughly with excess water.
• Incompatibilities: Avoid strong oxidizers and extremes of pH or temperature that can denature proteins. Do not mix with organic solvents.
• Special risks: Repeated freeze–thaw can cause aggregation and loss of activity; follow aliquoting practices.

First-aid overview (general)

• Skin/eye contact: Rinse with water for at least 15 minutes; remove contaminated clothing; seek medical advice if irritation persists.
• Ingestion/inhalation: Rinse mouth, move to fresh air, and seek medical advice if unwell.

Waste disposal

• Dispose of protein solutions and consumables according to institutional biosafety and chemical hygiene policies; if azide-containing, collect separately per local regulations.

Solvent Selection

Applicability

• As a protein biologic, this antibody is handled in aqueous buffers; organic solvent selection is generally not applicable.

Buffer compatibility (general guidance)

• Preferred: Neutral, isotonic buffers such as PBS or TBS, pH ~7.2–7.6.
• Additives sometimes used for stability (not item-specific): 10–50% glycerol for frozen storage, 0.01–0.05% nonionic detergents (Tween-20) in wash buffers to reduce nonspecific binding, carrier proteins (BSA) to minimize adsorption.
• Avoid: Strong acids/bases, high concentrations of chaotropes, and most organic solvents that denature proteins.

Comparison (general)

• PBS vs TBS: PBS may interact with phospho-sensitive assays; TBS can be preferred for alkaline phosphatase detection systems.
• Detergents: Tween-20 commonly used in immunoassays; higher concentrations can strip weak interactions.

Note: Actual formulation and compatible buffers for Ab133237 are not specified for this item; refer to the CoA/Spec Sheet. If performing specialized applications (e.g., conjugation), ensure buffer lacks amines, BSA, or azide as required by the chemistry.

Storage and Reconstitution

Item-specific conditions

• Storage Conditions: Store at -20°C. Avoid repeated freezing and thawing.
• Shipped In: Ice chest + Ice pads (cold chain).
• Reconstitution: Not specified for this item; refer to CoA/Spec Sheet.
• Buffer composition/preservatives: Not specified for this item; refer to CoA/Spec Sheet.

General handling guidance (advisory)

• Upon receipt: Inspect for container integrity and, if liquid, gentle mixing by inversion after equilibrating on ice. If lyophilized, briefly spin down before opening in a clean environment.
• Aliquoting: Prepare small working aliquots to prevent multiple freeze–thaw cycles. Use low-protein-binding tubes.
• Freezing: For long-term storage at -20°C, many antibodies benefit from addition of sterile glycerol to 40–50% v/v (general guidance; not item-specific). Avoid frost-free freezers.
• Thawing: Thaw on ice; mix gently. Do not vortex vigorously.
• Short-term storage: At 2–8°C for days to a few weeks is common for many formulations, but the allowed duration for Ab133237 is not specified; defer to CoA/Spec Sheet.

Stability notes

• Repeated freeze–thaw can lead to aggregation and reduced activity; plan aliquots accordingly.
• If sterility is required, work in a biosafety cabinet and use sterile, filtered buffers.

Always consult the product CoA/Spec Sheet and SDS for authoritative, lot-specific instructions.

Structure and Identity

Product type: mouse monoclonal antibody raised against the UBE2E3 protein (Ubiquitin-conjugating enzyme E2 E3). This is a protein biologic, not a small molecule.

Item-specific identifiers

• SKU: Ab133237
• Product Name: UBE2E3 Mouse mAb
• Category: Antibody (research use only)

Molecular identifiers (item-specific)

• Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES/InChI/InChIKey: Not applicable for proteins/antibodies.
• Clone, isotype, light chain type, and host species: Host species indicated as mouse; clone/isotype not specified for this item; refer to CoA/Spec Sheet.

Structural features (general, literature)

• Typical IgG monoclonal architecture with two heavy and two light chains forming a Y-shaped molecule; contains variable domains (VH/VL) for antigen binding and constant domains (CH1–CH3, CL) mediating Fc functions.
• Approximate mass for a full-length IgG is typically ~150 kDa (literature, varies by isotype and glycosylation).
• Post-translational modifications may include conserved Fc N-glycosylation; exact glycoform is lot- and process-dependent (not specified for this item).

Target antigen overview (general)

• UBE2E3 is a ubiquitin-conjugating enzyme (E2) that transfers activated ubiquitin from E1 to substrates in conjunction with E3 ligases; predominantly cytosolic/nuclear localization depending on cell type (literature).

Synthetic Utility

Not applicable. This is a mouse monoclonal antibody intended for biochemical detection of the UBE2E3 protein. It is not a building block or reagent for chemical synthesis. For practical experimental guidance, see Reaction & Applications and Application Protocols.

Target Specificity

Item-specific details

• Antigen/epitope: UBE2E3 (ubiquitin-conjugating enzyme E2 family). Exact immunogen/epitope is not specified for this item; refer to CoA/Spec Sheet.
• Species reactivity (tested/predicted): Not specified for this item; refer to CoA/Spec Sheet.
• Clone designation, isotype, and subclass: Not specified for this item; refer to CoA/Spec Sheet.
• Cross-reactivity: Not specified for this item; refer to CoA/Spec Sheet.

General background (literature)

• UBE2E3 is an E2 enzyme participating in the ubiquitination cascade. Antibodies to UBE2E3 typically recognize a conserved region of the protein; cross-species reactivity depends on sequence homology and epitope choice.

Recommendations

• Verify target size and isoforms for the species/sample by consulting UniProt/RefSeq prior to WB/IHC.
• Include positive controls (cells/tissues known to express UBE2E3) and negative controls (knockdown/knockout if available) to validate specificity in your system.

Domande frequenti

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 (UBE2E3 Mouse mAb); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.