Recombinant BrdU Antibody - Anticorpo primario, specific to BrdU

Cat. No.: Ab210601

GRADE & PURITY Animal Free, Carrier Free, Recombinant, ExactAb™, Azide Free, Validated, High Performance, PBS Only, ≥95%(SDS-PAGE), 1.0 mg/mL

Application

ELISA, IF/ICC

Host species

Mouse

Reactive species

Indipendente dalla specie

Isotype

Mouse IgG1

Clone Number

P59D18

Conjugation

Unconjugated

Panoramica

Recombinant BrdU Antibody, clone P59D18, could be used for IF/ICC, ELISA and so on

Specifications

Product Name

Recombinant BrdU Antibody - Anticorpo primario, specific to BrdU

Sinonimi

Bromodeoxyuridine antibody | BrdU antibody | Anti-BrdU antibody | 5-bromo-2-deoxyuridine antibody | BrdU Monoclonal Antibody | Anti-BrdU/5-BrdU Antibody | Anti BrdU

Specifiche e purezza

Animal Free,Carrier Free,Recombinant,ExactAb™,Azide Free,Validated,High Performance,PBS Only,≥95%(SDS-PAGE),1.0 mg/mL

Meccanismi biochimici e fisiologici

The immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of tumor cells with the thymidine analogue BrdU and the subsequent

Specie ospite

Mouse

Specificità

BrdU

Immunogeno

Bromodeoxyuridine

Controllo positivo

IF/ICC: HeLa cells

Coniugazione

Unconjugated

Grado

Animal Free, Azide Free, Carrier Free, ExactAb™, High Performance, PBS Only, Recombinant, Validated

Tipo di catena leggera

lambda

SDS-PAGE

150 kDa

Metodo di purificazione

Protein A purified

Concentrazione

1.0 mg/mL

Product Line

biological_products

Stoccaggio e spedizione

Condizioni di conservazione di stoccaggio

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

Spedito in

Ice chest + Ice pads

Stabilità e conservazione

Store at 4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

Immagini

IF/ICC: Recombinant BrdU Antibody (Ab210601)

Recombinant BrdU Antibody (Ab210601) - IF/ICC
IF analysis of Recombinant BrdU Antibody (Green) in HeLa cells. The cells were fixed with ethanol for 15 minutes and subjected to acid hydrolysis using 2M HCl for 15 minutes at room temperature to denature the DNA. They were then stained Recombinant BrdU Antibody (Ab210601) at 1.0 μg/mL in blocking buffer overnight at 4℃ and then incubated with Goat Anti-Mouse IgG H&L (FITC) (Ab179003) at a dilution of 1/1000 for 1 hour at room temperature in the dark. Cells were counterstained with DAPI (Blue). Images were taken on the confocal laser scanning microscope.

Applicazione

Applicazione

Dilution info

IF/ICC

1.0 μg/mL

IF/ICC

1.0 μg/mL

Application Protocols

• Item-specific

  • Tested applications, recommended dilutions, incubation times, and positive controls are Not specified for this item; refer to CoA/Spec Sheet.

• General workflow notes (for planning only)

  • BrdU labeling: Pulse cells/tissues with BrdU under experimental conditions; fix according to matrix requirements (e.g., paraformaldehyde for cells, formalin for tissues).
  • DNA exposure: Denature (e.g., acid) or nick (e.g., DNase I) DNA to reveal BrdU epitopes; neutralize thoroughly before antibody incubation.
  • Antibody incubation: Perform in PBS/TBS with appropriate blocking. Optimize concentration empirically.
  • Detection: Use compatible secondary antibodies or directly labeled conjugates if you perform custom labeling. Avoid sodium azide with HRP systems.

These are general considerations and not product‑validated protocols.

Biological Roles

• Target biology (literature/general)

  • BrdU (5‑bromo‑2′‑deoxyuridine) is a thymidine analog that cells can incorporate into DNA during S‑phase via the salvage pathway. It substitutes for thymidine triphosphate (TTP), enabling labeling of replicating DNA.
  • Anti‑BrdU antibodies recognize the BrdU hapten once DNA strands are denatured or nicked to expose the base, allowing immunodetection of proliferating cells.
  • Applications span cell cycle studies, neurogenesis tracking, stem/progenitor proliferation assays, and DNA repair dynamics when combined with pulse‑chase designs.

• Item-specific note

  • This product is an anti‑BrdU antibody for research use only; no biological activity beyond binding BrdU is claimed in the product data.

Buffer Applications

This section focuses on practical buffer systems for BrdU immunodetection (general guidance). No item-specific buffer recipes beyond “PBS only” are provided.

• Item-specific (Product Data)

  • Supplied in PBS only; azide-free; carrier-free.

• Literature/General practices

  • DNA denaturation/nicking: Common approaches include 1–2 N HCl for 10–30 min at RT to denature DNA or DNase I treatment under controlled conditions; neutralize/rehydrate before antibody incubation.
  • Blocking buffers: Protein-based blockers (e.g., 1–5% serum or BSA) can reduce nonspecific binding; avoid adding carriers to the antibody stock if you intend to perform conjugation.
  • Wash buffers: PBS or TBS with 0.05% Tween‑20 is typical for stringent washing while preserving binding.
  • pH range: Maintain near-neutral pH (7.2–7.6) during antibody incubation; strictly follow optimized conditions for your matrix (cells vs. tissue sections).
  • Permeabilization: Saponin or Triton X‑100 at low percentages may be used before denaturation to facilitate reagent access.

Green Alternatives

While green chemistry is less central for a protein reagent, several choices can reduce environmental impact without compromising performance.

• Item-specific positives

  • Azide-free: Eliminates sodium azide, reducing hazardous preservative handling and waste.
  • Carrier-free: Minimizes additional protein excipients, simplifying waste streams.

• General greener practices (literature/general)

  • Right-size aliquots: Reduce freeze–thaw waste and product loss, lowering repeat shipments.
  • Cold-chain efficiency: Consolidate orders to optimize refrigerated shipments and packaging waste.
  • Aqueous systems: Maintain purely aqueous buffers (PBS/TBS) and avoid organic cosolvents.

• Trade-offs

  • Azide-free solutions require stricter aseptic handling to prevent microbial growth, which may modestly increase single-use plastics (filters, sterile vials). Balance by planning experiments to minimize disposables.

Pharmaceutical Uses

Not applicable. This recombinant anti‑BrdU antibody is designated for research use only and is not supplied with any pharmacopeial status or clinical formulation claims.

• Item-specific (Product Data)

  • Research Use Note: For research use only.

• General information

  • Anti‑BrdU antibodies are widely used in nonclinical analytical assays to quantify cell proliferation in vitro and in preclinical models. No therapeutic, diagnostic, or clinical application is claimed or implied for this item.

Physical Properties

• Item-specific (Product Data)

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Formulation: PBS only; azide-free; carrier-free (as supplied).
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

• Literature/General values for context (not item specifications)

  • Typical IgG MW: ~150 kDa for a full-length IgG; recombinant formats (scFv, Fab, Fc‑fusion) can vary widely (~25–150 kDa).
  • Solubility: Immunoglobulins are water-soluble in physiological buffers (e.g., PBS, pH ~7.2–7.4); stability is sensitive to pH extremes, surfactants, and high ionic strength.
  • Isoelectric point (pI): Commonly ~6–9 for monoclonal antibodies, sequence dependent.
  • Stability considerations: Freeze–thaw cycles can promote aggregation; gentle handling and aliquoting are recommended.

Quality and Grades

• Item-specific (Product Data) and implications

  • Recombinant: Produced from a defined expression system for batch-to-batch consistency and animal-origin–free sequence control.
  • Animal Free: Manufactured without animal-derived raw materials, reducing risk of adventitious agents and enhancing regulatory compatibility for certain workflows.
  • Carrier Free: No added protein stabilizers (e.g., BSA). Advantages: precise assay backgrounds, compatibility with conjugation/labeling. Consider aliquoting to mitigate adsorption losses.
  • Azide Free: No sodium azide preservative. Advantages: compatible with peroxidase-based assays and certain cell-associated workflows; requires aseptic technique to prevent contamination.
  • ExactAb™ / Validated / High Performance: Indicates enhanced lot-to-lot consistency, functional validation, and high assay performance (as per manufacturer program). Specific validation datasets are not provided here; refer to CoA/Spec Sheet.
  • PBS Only: Supplied in phosphate-buffered saline without additional excipients.

• Notes on what is and isn’t specified

  • Purity metrics (SDS-PAGE homogeneity, HCP/DNA, endotoxin), concentration, isotype, clonality, and host are Not specified for this item; refer to CoA/Spec Sheet.
  • For sensitive applications (flow cytometry, ICC/IHC, ELISA, IF), verify lot-specific validation and recommended working ranges on the CoA/Spec Sheet.

Reaction and Applications

• Item-specific (Product Data)

  • Intended use: Research use only antibody recognizing BrdU; azide-free, carrier-free; supplied in PBS. No manufacturer application list is specified for this item; refer to CoA/Spec Sheet.

• Literature/General applications for anti‑BrdU antibodies

  • Cell proliferation assays: Detection of BrdU incorporation into newly synthesized DNA to quantify S‑phase cells in cultured cells or tissue sections.
  • Immunocytochemistry/Immunohistochemistry: After BrdU pulse and fixation, DNA is typically denatured (e.g., 1–2 N HCl) or nicked enzymatically to expose BrdU, then probed with anti‑BrdU.
  • Flow cytometry: Post‑incorporation permeabilization and DNA denaturation/nicking followed by intracellular staining enables cell cycle profiling.
  • ELISA/HTS formats: Plate-based detection of BrdU-labeled DNA for proliferation or genotoxicity assays.

• Practical tips (general)

  • DNA exposure is critical: Optimize denaturation conditions to balance epitope exposure and cellular/tissue morphology.
  • Controls: Include no‑BrdU and isotype controls, plus a positive BrdU‑pulsed control population.
  • Conjugation: Carrier‑free, azide‑free format is suitable for custom labeling (e.g., HRP, fluorophores); maintain gentle conditions (pH 7–8.5) to preserve activity.

Reaction Conditions

Not applicable in the context of chemical synthesis. For this antibody, the relevant “conditions” are immunostaining/incubation parameters, which are application-specific and should be optimized empirically.

• General guidance (literature)
  • Incubations are typically performed at 4 °C overnight or at room temperature for 1–2 h in PBS/TBS. Exact times, concentrations, and temperatures must be optimized per sample type and are not specified for this item; refer to CoA/Spec Sheet and in‑house method development.
  • Critical step: DNA denaturation or nicking to expose BrdU prior to antibody binding.

Safety and Handling

• Item-specific (Product Data)

  • GHS classification / pictograms / H-statements / signal word: Not specified for this item; refer to SDS.
  • Storage: Store at −20 °C. Avoid repeated freezing and thawing.
  • Shipping: Shipped in an ice chest with ice pads.
  • Azide-free: No sodium azide preservative in the supplied material.
  • Research Use: For research use only.

• General laboratory safety guidance (literature/best practice)

  • PPE: Wear lab coat, appropriate gloves (e.g., nitrile), and safety glasses when handling.
  • Handling: Keep on ice during use; minimize time at ambient temperature. Avoid vigorous vortexing; mix by gentle inversion.
  • Incompatibilities: Avoid strong oxidants, extreme pH, and organic solvents that denature proteins; avoid microbial contamination.
  • First aid overview: Protein solutions are generally of low acute hazard; in case of contact, rinse with water. If inhaled as aerosol, move to fresh air. Seek medical attention if irritation persists. Always follow the SDS for authoritative instructions.
  • Waste: Dispose of according to institutional biosafety procedures for non-hazardous biologicals unless local rules dictate otherwise.

Solvent Selection

This product is a protein reagent, not a solvent. Selection considerations focus on compatible assay buffers rather than organic solvents.

• Item-specific (Product Data)

  • Supplied in PBS only; azide-free; carrier-free.

• Practical buffer selection (literature/general)

  • Working buffer: PBS or TBS at pH 7.2–7.6 is commonly used. Maintain isotonicity and moderate ionic strength to preserve binding.
  • Additives: If absolutely necessary in your workflow, small amounts of nonionic detergents (e.g., 0.01% Tween‑20) can reduce nonspecific binding; however, avoid altering the supplied material unless your assay requires it. For conjugation, keep carrier/detergent free.
  • Avoid: High concentrations of denaturants (urea, guanidinium), organic solvents, and extreme pH which can impair antigen binding.
  • DNA exposure for BrdU detection: Buffer choice must accommodate DNA denaturation (e.g., HCl treatment) or enzymatic digestion (e.g., DNase I) steps before antibody incubation.

Storage and Reconstitution

• Item-specific (Product Data)

  • Storage temperature: Store at −20 °C.
  • Handling: Avoid repeated freezing and thawing; aliquot upon first thaw to single‑use volumes.
  • Shipping: Shipped in an ice chest with ice pads to maintain cold chain.
  • Formulation as supplied: PBS only; azide‑free; carrier‑free.

• Practical guidance (general)

  • Aliquoting: Dispense into low‑bind, sterile microtubes. Keep on ice during handling; minimize headspace to reduce oxidation.
  • Freeze–thaw: One freeze–thaw can be tolerated by many antibodies, but multiple cycles promote aggregation and activity loss. Store aliquots at −20 °C (or −80 °C if long‑term), per your internal stability data.
  • Thawing: Thaw on ice; mix gently by inversion. Do not vortex vigorously.
  • Short-term storage: Keep at 2–8 °C for brief use (hours to a few days) to avoid repeated freezing.
  • Additives: Because the product is carrier‑free and azide‑free by design, avoid adding stabilizers or preservatives unless your application specifically requires them; any additions change the supplied grade.
  • Light sensitivity: Protect from prolonged light exposure if conjugated post‑purchase with fluorophores.

For definitive stability, concentration, and buffer composition details, consult the lot-specific CoA/Spec Sheet.

Structure and Identity

Overview: This is a recombinant antibody directed against the DNA base analog bromodeoxyuridine (BrdU), supplied azide‑free in PBS and designated for research use only.

• Item-specific (Product Data)

  • Product Type: Recombinant anti‑BrdU antibody (ExactAb™, Animal Free, Carrier Free, Azide Free, Validated, High Performance, PBS Only)
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Sequence/clonality/isotype: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES / InChI / InChIKey: Not applicable to proteins; not specified for this item.

• Literature/General information (for context)

  • Target: BrdU (5-bromo-2′-deoxyuridine), a thymidine analog incorporated into DNA during S-phase.
  • Antibody architecture (typical for IgG): Two heavy and two light chains forming a Y‑shaped immunoglobulin with variable domains that recognize the BrdU hapten when exposed in DNA after denaturation or nuclease treatment.
  • Epitope nature: Small-molecule (hapten) covalently incorporated into DNA; detection usually requires DNA strand denaturation to expose BrdU.

Synthetic Utility

Not applicable. As a biological reagent (antibody), this product is not used as a synthetic building block or reagent in organic synthesis.

• For related utility, see “Reaction & Applications” for immunodetection workflows and “Buffer Applications” for assay buffers.

Target Specificity

• Item-specific (from Product Name)

  • Antigen/Target: BrdU (5‑bromo‑2′‑deoxyuridine), a thymidine analog incorporated into DNA during replication.

• Not specified for this item; refer to CoA/Spec Sheet for the following details

  • Clone designation and clonality (monoclonal/polyclonal)
  • Host species and expression system
  • Isotype/subclass and light chain type
  • Confirmed species/tissue reactivity and cross-reactivity profile
  • Affinity/avidity metrics and epitope mapping (if available)

Note: BrdU is a small-molecule hapten; species reactivity is typically not applicable in the conventional sense but matrix compatibility (cells, tissues) depends on processing and denaturation protocol.