Recombinant BAP1 Antibody - Primary antibody, specific to BAP1

Cat. No.: Ab325693

GRADE & PURITY KO Validation

Application

WB

Host species

Rabbit

Reactive species

Human, Mouse, Rat

Isotype

Rabbit IgG

Conjugation

Unconjugated

Specifications

Product Name

Recombinant BAP1 Antibody - Primary antibody, specific to BAP1

Spécifications et pureté

KO Validation

Espèce hôte

Rabbit

Spécificité

BAP1

Immunogène

Recombinant protein of human BAP1

Conjugation

Unconjugated

Grade

KO Validation

Méthode de purification

Protein A purified

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

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

Expédié en

Ice chest + Ice pads

Stabilité et stockage

Store at -20°C for one year.

Application Protocols

• Item-specific tested applications, recommended dilutions, species reactivity, positive controls, and detection systems: Not specified for this item; refer to CoA/Spec Sheet.

• General workflow outlines (literature/general; not specific to this SKU):

  • Western blot: Prepare lysates with protease inhibitors; quantify protein; run SDS-PAGE; transfer; block (e.g., 5% BSA); incubate with primary antibody; wash; incubate with species-appropriate secondary; detect by chemiluminescence or fluorescence.
  • Immunoprecipitation: Lyse cells under non-denaturing conditions; pre-clear; incubate lysate with antibody; capture with Protein A/G resin; wash; elute and analyze by WB or MS.
  • Immunofluorescence/IHC: Fix (e.g., PFA), permeabilize as needed, block, incubate with primary, wash, detect with labeled secondary; optimize antigen retrieval for FFPE tissues.

• Important note: Because application details for Ab325693 are not provided, users should perform preliminary titrations and control experiments, especially when working with BAP1 KO/WT pairs for specificity assessment.

Biological Roles

• Target overview (literature/general):

  • BAP1 (BRCA1-associated protein 1) is a nuclear deubiquitinase (DUB) of the UCH family. It functions within multiprotein complexes regulating chromatin dynamics, histone H2A deubiquitination (particularly H2AK119ub1), transcription, cell cycle progression, and DNA damage responses.
  • BAP1 forms complexes with ASXL proteins to constitute the PR-DUB complex, counteracting Polycomb-mediated gene silencing. It also participates in regulation of protein stability via deubiquitination of specific substrates.
  • Loss-of-function mutations or deletions in BAP1 are frequently observed in certain cancers in research literature; these studies leverage BAP1 antibodies for expression profiling and mechanistic assays. No clinical/diagnostic claims are made here; for research use only.

• Experimental implications (general):

  • Subcellular localization: Predominantly nuclear; extraction buffers that preserve nuclear proteins (e.g., mild detergent plus salt) are typically used.
  • Post-translational modifications of BAP1 and its association with chromatin can influence epitope accessibility; antigen retrieval and fixation conditions should be empirically optimized.

Buffer Applications

• Applicability:

  • As an antibody reagent, buffer selection directly impacts performance in immunoassays. Item-specific buffer composition is not specified; consult the CoA/Spec Sheet.

• General buffer guidance for antibody workflows (literature/general):

  • Dilution buffers: PBS or TBS at pH 7.2–7.6; include 0.05–0.1% Tween-20 for wash steps to reduce nonspecific binding. Add 0.1–1% BSA or casein as a carrier/blocker where appropriate.
  • Blocking solutions: 3–5% BSA, casein, or species-appropriate serum; choose based on detection system to avoid cross-reactivity.
  • Antigen retrieval (IHC): Heat-induced retrieval in citrate (pH ~6) or Tris-EDTA (pH ~9) buffers; optimal buffer depends on tissue and fixation.
  • Storage/dilution considerations: For working stocks used over several days, include 0.02–0.05% sodium azide when compatible with downstream assays. For enzyme-based detection systems, avoid azide.
  • Filtration/sterility: 0.22 µm filtration for long-term working solutions if preservative-free; use low-protein-binding consumables.

Green Alternatives

• Context:

  • Environmental considerations for antibody use focus on buffer components, blocking agents, and wash solutions rather than organic solvent selection.

• Greener practice suggestions (literature/general):

  • Replace methanol-based fixation or deparaffinization steps with lower-toxicity alternatives where compatible (e.g., ethanol gradients; commercial xylene substitutes), understanding performance trade-offs.
  • Use phosphate- or Tris-based buffers prepared with high-purity water; avoid azide when downstream enzymatic assays or live-cell work are anticipated. If azide-free formulations are required, maintain sterility via filtration and small aliquots to reduce bioburden.
  • Minimize single-use plastics by employing reusable staining containers and calibrated wash bottles; select low-binding, recyclable plastics when feasible.
  • Concentrate wash volumes and adopt microplate/slide chambers that reduce reagent consumption.

• Trade-offs:

  • Preservative-free buffers reduce hazardous waste but shorten shelf life; balance stability vs. hazard profile based on experimental cadence.
  • Xylene substitutes and aqueous clearing agents may alter staining quality; validate against your assay controls.

Pharmaceutical Uses

• Applicability to this product:

  • This is a research-grade recombinant antibody. It is not intended for therapeutic, diagnostic, or clinical use.

• General notes (literature/general):

  • Research antibodies can be used in pharmaceutical R&D for target validation, mechanism-of-action studies, biomarker discovery, and QC analytics in process development. Such uses require internal qualification for specificity, sensitivity, and matrix effects.
  • Pharmacopeial/compendial status: Not applicable; no pharmacopeial monograph is implied for this item.
  • If antibodies are used as analytical reagents in regulated environments, users should establish method suitability (specificity, linearity, precision) and assess stability under intended storage conditions. Item-specific validation data are not provided here.

Physical Properties

• Item-specific (from Product Data):

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

• General properties for recombinant antibodies (literature/general):

  • Approximate molecular mass: IgG antibodies typically ~150 kDa; fragments (Fab, scFv) are smaller. Exact value depends on construct and glycosylation.
  • Physical state: Supplied in aqueous buffer or lyophilized; this item’s formulation is not specified.
  • Solubility: Readily soluble in common biological buffers (PBS, TBS) at working concentrations; stability influenced by pH (~7–8), ionic strength, and excipients (e.g., glycerol, BSA, sugars). Specific buffer for this item is not specified.
  • pI/logP/RI/BP/MP: Not applicable/typically not reported for macromolecular biologics in catalog specifications.

• Practical notes (general):

  • Avoid foaming and vigorous vortexing; proteins can denature at air–liquid interfaces.
  • Minimize exposure to elevated temperature; antibodies gradually lose activity with repeated freeze–thaw.

Quality and Grades

• Item-specific (from Product Data):

  • Grade/Purity: KO Validation.

• Interpretation and implications (literature/general):

  • KO (knockout) validation indicates the antibody’s target specificity was evaluated by testing in a biological context lacking the target gene/protein. A high-quality KO-validated antibody should show loss/absence of the specific signal in KO/knockdown samples compared to wild-type. While the product carries the KO Validation designation, exact datasets, sample types (cell line/tissue), and supported applications are not specified for this item; consult the CoA/Spec Sheet for validation images and conditions.
  • Recombinant production generally affords lot-to-lot consistency compared to hybridoma-derived monoclonals or polyclonals; however, clonality, isotype, concentration, and buffer composition are not specified here and should be confirmed on the accompanying documentation.
  • QC elements commonly assessed for antibodies include concentration by A280, purity by SDS-PAGE, endotoxin (for cell-based assays), and functional validation in specific applications (WB, IHC, IF, IP). These specific metrics are not specified for this item.

Reaction and Applications

• Applicability to this product:

  • This is a biological affinity reagent rather than a chemical reactant. No chemical reactions or named synthetic transformations apply.

• Research application context (literature/general; not manufacturer-validated for this SKU unless stated):

  • Antibodies to BAP1 are commonly used for:
    • Western blotting to detect BAP1 expression and assess KO/knockdown efficiency.
    • Immunoprecipitation or chromatin immunoprecipitation (ChIP) to study BAP1-associated complexes and chromatin functions.
    • Immunofluorescence/IHC to determine subcellular localization and tissue distribution of BAP1.
  • Epitope retrieval, fixation methods, and lysis conditions must be optimized for BAP1, a nuclear deubiquitinase associated with chromatin. Conditions that preserve protein–protein interactions (mild detergents, DNase for viscosity) may be preferred for IP.
  • For this item, specific tested applications, species reactivity, and working dilutions are not specified; consult CoA/Spec Sheet and perform in-lab optimization.

Reaction Conditions

• Not applicable to this product:

  • There are no chemical reaction conditions associated with the intrinsic use of this antibody.

• Assay condition considerations (literature/general):

  • For immunoassays, temperature (room temperature vs. 4°C incubations), incubation times, and buffer composition strongly influence affinity and background. Start with gentle conditions and optimize empirically.
  • Detergent choice (e.g., 0.1% Triton X-100, 0.1% Tween-20) and salt concentrations modulate epitope accessibility and specificity; nuclear proteins like BAP1 may benefit from optimized extraction protocols.
  • Specific, item-tested conditions are not provided; consult CoA/Spec Sheet.

Safety and Handling

• Item-specific (from Product Data):

  • Storage: Store at -20°C. Avoid repeated freezing and thawing.
  • Shipping: Ice chest + ice pads (cold pack shipment).
  • GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.

• General guidance (literature/general):

  • Recombinant antibodies are generally considered non-hazardous proteins but may be supplied in buffers containing preservatives (e.g., sodium azide) or glycerol; check the SDS/label for specific hazards.
  • PPE: Laboratory coat, nitrile gloves, and safety glasses. Handle using aseptic technique to prevent microbial contamination.
  • Incompatibilities: Avoid strong oxidizers and proteases; maintain recommended pH to prevent denaturation/aggregation. Do not mix with reducing agents if disulfide integrity is required.
  • First aid (overview; defer to SDS): Skin/eye contact—rinse with water. Inhalation—move to fresh air. Ingestion—not intended; rinse mouth. Seek medical attention if irritation persists.
  • Special risks: Repeated freeze–thaw cycles can cause aggregation and loss of activity; aliquot immediately upon receipt. Use low-protein-binding plasticware to minimize adsorption losses.

Solvent Selection

• Applicability to this product:

  • As a recombinant antibody supplied in aqueous buffer or lyophilized form, classical organic solvent selection is not typically applicable. Work instead with biological buffers compatible with immunoassays.

• General guidance for antibody buffers (literature/general):

  • Preferred diluents: PBS or TBS at pH 7.2–7.6 with optional 0.05–0.1% Tween-20 for washing steps in immunoassays.
  • Stabilizers: 10–50% glycerol for freeze–thaw protection at -20°C; carrier proteins (e.g., 0.1–1% BSA) can improve stability and reduce nonspecific binding. Specific formulation for this item is not specified.
  • Avoid: High concentrations of denaturants (urea, guanidinium), strong detergents (SDS), extreme pH (<6 or >8.5) unless required by a specific protocol, as these can compromise antigen binding.
  • Filtration: Use 0.22 µm low-protein-binding filters when sterility is required; avoid repeated filtration to minimize loss by adsorption.

Storage and Reconstitution

• Item-specific (from Product Data):

  • Storage conditions: Store at -20°C. Avoid repeated freezing and thawing.
  • Shipping: Ice chest + ice pads (cold pack shipment).
  • Appearance, concentration, formulation buffer, and preservative: Not specified for this item; refer to CoA/Spec Sheet.

• Practical guidance (literature/general):

  • Upon receipt, briefly centrifuge the vial to collect contents. If liquid, aliquot into low-protein-binding tubes sized for single use to prevent freeze–thaw.
  • If lyophilized (not specified for this item), reconstitute with sterile buffer (e.g., PBS) to the desired concentration; allow to dissolve gently on ice without vigorous mixing. Record final concentration and buffer.
  • For -20°C storage, inclusion of 30–50% glycerol in the storage buffer is commonly used to permit storage without freezing; verify compatibility with your applications and the product’s formulation.
  • Short-term storage (≤1 week) at 2–8°C may be acceptable for working aliquots; monitor for turbidity or precipitation.
  • Do not store in frost-free freezers that cycle temperatures. Avoid repeated freeze–thaw as specified.
  • Always refer to the SDS and CoA/Spec Sheet for item-specific buffer composition, stability, and reconstitution recommendations.

• Research use note: For research use only.

Structure and Identity

• Item-specific (from Product Data):

  • SKU: Ab325693
  • Product name: Recombinant BAP1 Antibody
  • Grade/Purity: KO Validation
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • CAS/CID/InChIKey/SMILES: Not applicable to protein biologics and not specified for this item.

• General characterization (literature/general):

  • This product is a recombinant antibody directed against BAP1 (BRCA1-associated protein 1), a deubiquitinase implicated in chromatin regulation and tumor suppression.
  • Antibody type/isotype, clonality, host species, and epitope are not specified for this item; consult the CoA/Spec Sheet.
  • Typical 2D structural description for antibodies: Y‑shaped immunoglobulin with two identical heavy chains and two identical light chains linked by disulfide bonds; variable domains confer antigen binding. Recombinant production often ensures sequence-defined variable regions and batch-to-batch consistency.
  • Post-translational features (glycosylation, Fc modifications) can affect Fc receptor interactions and stability; details are not specified for this item.

Synthetic Utility

• Not applicable to this product:

  • Recombinant antibodies are biological affinity reagents and are not used as building blocks or reagents in organic synthesis.

• Guidance (literature/general):

  • For chemical conjugation (e.g., labeling), antibodies can be derivatized at lysine ε-amines or reduced interchain cysteines using NHS esters or maleimide chemistries, respectively. If conjugation is planned, confirm buffer composition (amine- and thiol-free) and desalt as needed.
  • No synthetic transformations or retrosynthetic roles pertain to the antibody itself.

Target Specificity

• Item-specific (from Product Data):
  • Target/Antigen: BAP1 (BRCA1-associated protein 1).
  • Additional details (epitope, species reactivity, clonality, isotype, cross-reactivity): Not specified for this item; refer to CoA/Spec Sheet.
  • KO Validation grade indicates evaluation against BAP1 knockout context; specific datasets and applications are not provided here.

Foire aux questions

What does KO Validation mean?

KO 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.