PCCA Mouse mAb - Primary antibody, specific to PCCA

Cat. No.: Ab325651

GRADE & PURITY KD Validation

Application

IF/ICC, WB

Host species

Mouse

Reactive species

Human, Mouse, Rat

Isotype

Mouse IgG1

Conjugation

Unconjugated

Specifications

Product Name

PCCA Mouse mAb - Primary antibody, specific to PCCA

Specifications & Purity

KD Validation

Host species

Mouse

Specificity

PCCA

Immunogen

Recombinant protein of human PCCA

Conjugation

Unconjugated

Grade

KD Validation

Purification Method

Protein A/G purified

Product Line

Biological Products

Storage and Shipping

Storage

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

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20°C for one year.

Application Protocols

Specific, tested application protocols and dilutions for Ab325651 are not provided in the product data. Please consult the CoA/Spec Sheet for lot‑specific recommendations.

General starting points (for mouse mAbs; not item-specific):

• Western blot (WB): Start at 1:1,000 in TBST with 3–5% BSA; incubate 1 h at room temperature or overnight at 4°C. Validate band size and include KD/KO control if available.
• Immunofluorescence/ICC: Start at 1:200 in blocking buffer; fix with 4% PFA, permeabilize with 0.1% Triton X‑100 if epitope is intracellular; incubate 1 h at room temp or overnight at 4°C.
• IHC: Begin at 1:200; perform heat‑induced antigen retrieval (e.g., citrate pH 6.0 or Tris‑EDTA pH 9.0); optimize blocking and detection chemistry.
• IP: 1–5 µg antibody per mg lysate protein; couple to Protein G/A matrix suited to the mAb’s isotype (isotype not specified here).

Controls and validation:

• Include no‑primary, isotype, and secondary‑only controls.
• For KD‑validated antibodies, confirm signal loss upon target knockdown in your system.

These are generic guidelines and should be titrated for sample type, detection system, and instrument sensitivity.

Biological Roles

Target context (PCCA; literature/general biology):

• Propionyl‑CoA carboxylase (PCC) is a mitochondrial, biotin‑dependent enzyme that converts propionyl‑CoA to D‑methylmalonyl‑CoA, an anaplerotic step linking the catabolism of odd‑chain fatty acids, cholesterol side‑chains, and certain amino acids (isoleucine, valine, threonine, methionine) to the tricarboxylic acid (TCA) cycle via succinyl‑CoA.
• PCC is a heterododecamer composed of alpha (PCCA) and beta (PCCB) subunits. The PCCA subunit harbors the biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) domains, including the conserved lysine to which biotin is covalently attached. PCCB contains the carboxyltransferase activity; together they perform ATP‑dependent carboxylation using bicarbonate as the CO2 source.
• Subcellular localization: Predominantly mitochondrial matrix; import requires an N‑terminal transit peptide on the precursor polypeptide (literature).
• Regulation: Availability of biotin, substrate levels (propionyl‑CoA), and mitochondrial health influence flux through this pathway; post‑translational modifications on mitochondrial enzymes can modulate activity (literature context).

Relevance for research use:

• Monitoring PCCA protein levels supports studies of mitochondrial metabolism, nutrient utilization, and isotope‑tracing experiments through the anaplerotic node.
• Genetic perturbation (KD/KO) or nutrient challenges can be paired with this antibody to assess pathway adaptation at the protein level.

Note: The above summarizes general biology; it does not assert specific performance characteristics of this antibody lot.

Buffer Applications

This product is an antibody reagent rather than a buffering compound. Traditional buffer preparation guidance (pKa, buffering ranges) is not applicable.

Practical notes (general):

• Antibody working solutions are typically prepared in PBS or TBS with blocking proteins (e.g., 1–5% BSA or milk for WB) and optional 0.05% Tween‑20 for washes.
• For IHC/IF, include carrier proteins and detergents compatible with fixation/permeabilization methods.

Item-specific buffer composition, recommended diluents, and pH for this SKU are not specified; please refer to the CoA/Spec Sheet.

Green Alternatives

While “green chemistry” frameworks mainly target solvents and synthetic reagents, sustainability considerations for antibodies focus on formulation, handling, and waste:

Greener handling practices (general suggestions):

• Aliquot to minimize freeze–thaw cycles, thereby reducing product loss and repeat shipments.
• Use reusable cold packs and insulated shippers; consolidate orders to reduce packaging waste.
• Employ phosphate‑buffered saline (PBS) without azide when compatible with your application and disposal requirements, as azide complicates wastewater handling. Preservatives for this SKU are not specified; check CoA/Spec Sheet before buffer exchange.

Comparison of formulation options (general; not item-specific):

• With sodium azide: Improved microbial stability; requires hazardous waste stream and is incompatible with HRP‑based detection.
• Azide‑free: Broader assay compatibility; may require sterile technique and small aliquots or 0.02% ProClin‑type preservatives.
• 50% glycerol at −20°C: Enables stable, frost‑free storage without freezing; increases viscosity and carbon footprint but reduces freeze–thaw damage.

End-of-life:

• Segregate antibody and protein‑contaminated consumables as biological laboratory waste. Rinse and recycle outer packaging where permitted.

These are general considerations; for this SKU, formulation details are not specified—consult the CoA/Spec Sheet.

Pharmaceutical Uses

This product is designated For research use only and is not intended for diagnostic or therapeutic applications.

General formulation context (non‑clinical, laboratory use):

• Antibody reagents may serve as analytical controls or process development tools in biomanufacturing R&D (e.g., monitoring target expression in cell lines or validating knockdown efficacy during assay development). These uses are distinct from clinical diagnostics and require internal qualification.
• No pharmacopeial monograph or cGMP release criteria are provided for this SKU. Endotoxin levels, bioburden, sterile filtration status, and excipient composition are not specified and should not be presumed to meet clinical standards.

Any clinical, diagnostic, or therapeutic deployment would require separate validation, regulatory clearance, and manufacturing controls—outside the scope of this product.

Physical Properties

Item-specific physicochemical specifications (concentration, buffer composition, preservative, pH, protein content, extinction coefficient) are not listed for this SKU.

• Appearance/form: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration: Not specified for this item; refer to CoA/Spec Sheet.
• Buffer: Not specified for this item; refer to CoA/Spec Sheet (common antibody buffers are PBS or Tris with stabilizers; general information only).
• Preservatives: Not specified for this item; refer to CoA/Spec Sheet.
• pH: Not specified for this item; refer to CoA/Spec Sheet.
• Osmolality: Not specified for this item; refer to CoA/Spec Sheet.
• Refractive index, density, BP/MP, logP: Not applicable for protein solutions in a meaningful way.

Literature/general values for IgG (context only, not item-specific specs):

• Approximate molecular mass: ~150 kDa for a full-length IgG (varies with isotype and glycosylation).
• Isoelectric point (pI): Typically ~6–9 depending on isotype and sequence.
• Extinction coefficient (ε 280 nm): Often ~1.3–1.5 mL·mg⁻¹·cm⁻¹ for many IgGs; exact value depends on amino acid composition.
• Solubility: Generally highly soluble in aqueous buffers at neutral pH; sensitive to aggregation at extremes of pH, elevated temperature, or repeated freeze–thaw cycles.

Please consult the product’s CoA/Spec Sheet for definitive, lot-specific physical parameters.

Quality & Grades

• Grade/Purity (as listed): KD Validation.

Interpretation and implications (general guidance):

• KD (knockdown) validation indicates that antibody specificity was assessed by reducing target gene expression (e.g., siRNA/shRNA) and demonstrating a corresponding loss or reduction of the detected signal in the relevant assay. This strengthens confidence that the observed band/spot/staining is attributable to the intended target rather than off‑target binding.
• KD validation complements conventional controls (isotype control, no‑primary control, peptide competition when applicable) and is particularly meaningful for Western blot or immunofluorescence where target modulation can be quantified.

What is specific vs. general:

• Specific to this item: The catalog lists “KD Validation,” indicating the vendor recognizes knockdown-based verification for this antibody class or lot. Precise details (cell line, transfection method, KD efficiency, assay format, exposure settings) are not specified for this item; refer to CoA/Spec Sheet.
• Not specified: Purity by SDS‑PAGE, endotoxin level, host species, isotype/subclass, clonality metadata (clone ID), and cross‑reactivity. These may affect assay selection and should be checked on the lot documentation.

Practical takeaways:

• Even with KD validation, users should run appropriate in‑assay controls and confirm expected molecular weight and localization for PCCA in their system.
• For quantitative work, verify linear dynamic range and signal proportionality versus protein load or target modulation.
• Research Use Only (per product data); not for diagnostic or therapeutic use.

Reaction & Applications

Classical organic reaction chemistry is not applicable to an antibody reagent. Instead, applications are immunoassay‑focused.

Potential research applications (general guidance; item-specific validations are not provided):

• Western blotting (WB) to detect PCCA at its expected molecular weight in cell or tissue lysates.
• Immunofluorescence (IF) or immunocytochemistry (ICC) to visualize mitochondrial PCCA localization (general cellular context for the target; verify protocol compatibility).
• Immunohistochemistry (IHC) on fixed tissues, using appropriate antigen retrieval optimized for the epitope (parameters not specified for this item).
• Immunoprecipitation (IP) to enrich PCCA‑containing complexes for downstream mass spectrometry or activity studies.

Notes:

• Specific tested applications, recommended dilutions, and positive controls are not specified for this item; refer to CoA/Spec Sheet.
• Employ appropriate negative/positive controls and, where feasible, genetic perturbation (KD/KO) to confirm specificity (aligned with KD Validation grade).
• Optimize antibody concentration, incubation time, and blocking conditions empirically for your sample type.

Reaction Conditions

Classical reaction conditions (solvents, catalysts, temperatures) do not apply to an antibody reagent. For immunoassays, conditions are operational rather than chemical.

General operational conditions (not item-specific):

• Western blot: Block with 3–5% BSA or milk; incubate primary antibody typically 1–16 h at 4–25°C; optimize dilutions empirically. Wash with TBS‑T or PBS‑T.
• IF/ICC: Fixation (e.g., 4% PFA), permeabilization (e.g., 0.1–0.5% Triton X‑100) as appropriate; primary incubation 1–16 h; protect fluorophores from light.
• IHC: Antigen retrieval (citrate or EDTA buffers) and blocking steps tailored to tissue and fixation; detection via HRP/DAB or fluorescence.

For this SKU, tested applications, recommended dilutions, incubation times, and retrieval conditions are not specified. Users should begin with standard conditions for mouse mAbs and titrate to optimal signal‑to‑noise in their system, incorporating KD/KO or peptide‑competition controls where applicable.

Safety & Handling

Safety details specific to this SKU are not provided in the product data. Always consult the product SDS for authoritative information.

• GHS classification, signal word, pictograms, H‑statements: Not specified for this item; refer to SDS.
• General nature of hazard: Antibodies are protein biologics in aqueous buffer; they are typically handled at Biosafety Level 1 unless otherwise stated. Treat as a laboratory reagent; avoid ingestion, inhalation of aerosols, and skin/eye contact.

Good laboratory practice (GLP) recommendations:

• PPE: Lab coat, nitrile gloves, and safety glasses.
• Handling: Work on ice or at 2–8°C during manipulations to maintain stability. Minimize time at ambient temperature. Avoid generating aerosols.
• Hygiene: Wash hands after handling; do not eat/drink in lab areas.

Storage incompatibilities and stability considerations:

• Do not mix with oxidizing agents or proteases. Avoid metal contamination if the antibody is conjugated to metals (not specified for this item).
• Critical: Avoid repeated freeze–thaw cycles (per product data) to prevent denaturation and aggregation.

First aid (overview; defer to SDS):

• Skin/eye contact: Rinse with water for several minutes; remove contaminated clothing. Seek medical advice if irritation persists.
• Inhalation: Move to fresh air; seek medical attention if symptoms occur.
• Ingestion: Rinse mouth; seek medical attention.

Waste disposal:

• Dispose of unused material and contaminated consumables according to institutional biosafety procedures and local regulations for non-hazardous biological reagents.

Solvent Selection

This product is an antibody preparation intended for use in aqueous buffers (e.g., PBS or Tris-based). Classical organic solvent selection guidance for small molecules is not applicable.

• Working medium: Use physiological ionic strength buffers at neutral pH unless otherwise validated for your assay. High percentages of organic solvents can denature antibodies.
• Additives: Low concentrations of non‑ionic detergents (e.g., 0.05% Tween‑20) may be used in wash buffers to reduce nonspecific binding in immunoassays (general guidance). Specific formulation for this SKU is not provided; consult the CoA/Spec Sheet before adding stabilizers or detergents.

If specialized applications require solvent exposure (e.g., permeabilization):

• Prefer mild, brief treatments (e.g., 0.1–0.5% Triton X‑100 for fixed cells) validated for antibody integrity.
• Avoid DMSO or alcohols in the primary antibody solution unless validated, as they can disrupt binding affinity.

For practical guidance, see Application Protocols (not specified here) and Storage & Reconstitution for handling considerations.

Storage & Reconstitution

• Storage conditions (from product data): Store at −20°C. Avoid repeated freezing and thawing.
• Shipping: Ice chest + ice pads (per product data).

General handling and reconstitution guidance (not item-specific):

• Upon receipt, briefly centrifuge the vial to collect contents, keep on ice while setting up aliquots.
• If provided lyophilized: Reconstitute with sterile, antibody‑grade water or buffer to the recommended concentration (not specified here; refer to CoA/Spec Sheet). Gently invert; avoid vigorous vortexing.
• If provided in solution: Prepare single‑use aliquots in low‑bind tubes sized for your most common assay volumes to eliminate freeze–thaw cycles.
• For storage at −20°C without freezing, consider adding sterile glycerol to a final concentration of 50% v/v (general practice) if compatible with your downstream application; verify that the original formulation permits glycerol addition.
• Short‑term storage: 2–8°C for up to a few days during active use is typically acceptable for many antibodies; minimize time at room temperature.
• Light protection: If fluorophore‑conjugated (not specified for this item), protect from light.

Stability notes:

• Avoid microbial contamination; use sterile technique. Preservatives (e.g., sodium azide) are not specified for this SKU—check CoA before buffer exchange or long‑term strategy.

Always consult the CoA/Spec Sheet for lot‑specific concentration, formulation, and any reconstitution instructions.

Structure & Identity

PCCA Mouse mAb (SKU: Ab325651) is a mouse monoclonal antibody raised against the propionyl‑CoA carboxylase alpha subunit (PCCA). Item-specific molecular identifiers customary for small molecules (CAS, SMILES, InChIKey) do not apply to antibodies and are not provided for this product.

• Product type: Mouse monoclonal antibody (isotype, clone, and host details: Not specified for this item; refer to CoA/Spec Sheet.)
• Target: PCCA protein (propionyl‑CoA carboxylase alpha subunit) — target specificity details for this lot are not provided here.
• Molecular formula: Not applicable to proteins in the conventional sense.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Typical IgG MW ~150 kDa (literature, varies with glycosylation and isotype).
• Sequence/epitope: Not specified for this item; refer to CoA/Spec Sheet.
• Conjugation: Not specified for this item; refer to CoA/Spec Sheet.

2D/structural description (general for antibodies):

• Composed of two heavy chains and two light chains forming a Y‑shaped immunoglobulin with variable regions at the Fab arms responsible for antigen binding and an Fc region mediating effector/interactions. Disulfide bonds stabilize inter- and intrachain architecture. If unconjugated, no additional reporter moieties are present; if conjugated, a label would be covalently attached to lysines or engineered tags (not specified here).

Notes:

• Research Use Only (per product data).

Synthetic Utility

Not applicable. This is a biological antibody reagent rather than a synthetic building block or catalyst.

For users interested in conjugation chemistry (general guidance):

• Antibodies can be derivatized via lysine (NHS‑ester) or cysteine (maleimide) chemistry to attach fluorophores, biotin, or enzymes for detection. The conjugation status for this SKU is not specified; if conjugation is desired, verify buffer composition (amine‑containing buffers and azide can interfere with certain chemistries) and perform buffer exchange prior to labeling.
• Maintain mild conditions (pH 7.2–7.5 for NHS esters; pH 6.5–7.0 for maleimides) and control degree of labeling to preserve antigen binding.

These are general considerations and do not reflect item‑specific specifications.

Target Specificity

Item-specific target details are not provided.

• Antigen: Not specified for this item; refer to CoA/Spec Sheet. (Product name suggests PCCA as the intended target, but definitive antigen identity, immunogen region/peptide, and species reactivity are not listed here.)
• Clone ID and isotype: Not specified for this item; refer to CoA/Spec Sheet.
• Species reactivity and cross‑reactivity: Not specified for this item; refer to CoA/Spec Sheet.
• Tested applications and validation data: Not specified for this item; refer to CoA/Spec Sheet.

General expectations for a PCCA‑directed mouse mAb (context only):

• Should recognize the PCCA protein near its expected molecular weight in WB and show mitochondrial patterning by IF in PCCA‑expressing cells, with signal reduced upon PCCA knockdown (aligns with KD validation concept). Actual performance is assay‑ and lot‑dependent and must be verified empirically.

Frequently Asked Questions

What does KD Validation mean?

KD Validation 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.

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.