ICAD Antibody - Anticorpo primario, specific to ICAD, Rabbit IgG

Cat. No.: Ab109205

GRADE & PURITY Carrier Free, ExactAb™, Validated, High Performance, See COA

Application

IF/ICC, IHC, WB

Host species

Coniglio

Reactive species

Umano

Isotype

Rabbit IgG

Conjugation

Unconjugated

Panoramica

Rabbit anti Human ICAD Antibody, Polyclonal, could be used for WB, IHC and so on.

Application

WB: 0.01-2 μg/mL

IHC: 5-20 μg/mL

IF/ICC: 2.5 μg/mL

Protein Function

Inhibitor of the caspase-activated DNase (DFF40).

Specifications

Product Name

ICAD Antibody - Anticorpo primario, specific to ICAD, Rabbit IgG

Sinonimi

A330085O09Rik antibody | Caspase activated deoxyribonuclease inhibitor short form antibody | DFF 1 antibody | DFF 45 antibody | DFF alpha antibody | DFF-45 antibody | DFF1 antibody | DFF35 antibody | DFF45 antibody | DFFA antibody | Dffa DNA fragmentation

Specifiche e purezza

Carrier Free, ExactAb™, Validated, High Performance, See COA

Specie ospite

Coniglio

Specificità

ICAD

Immunogeno

Recombinant ICAD expressed in E.coli (AA 51-305).

Controllo positivo

WB: Jurkat, A431, Hep G2, K562 and MCF7 cell lysates. IHC: Human breast cancer and kidney tissues. IF/ICC: HeLa cells.

Coniugazione

Unconjugated

Grado

Carrier Free, ExactAb™, High Performance, Validated

Isotipo

Rabbit IgG

SDS-PAGE

150 kDa

Metodo di purificazione

Antigen affinity purified + Protein A purified

Concentrazione

See COA

Stoccaggio e spedizione

Forma

Liquid

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℃ short term (1-2 weeks). Store at -20℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Immagini

Western Blot: ICAD Antibody (Ab109205)

ICAD Antibody (Ab109205) - Western Blot
All lanes: ICAD Antibody (Ab109205) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) (Ab170144) at 1/20000 dilution

Predicted band size: 37 kDa
Observed band size: 41 kDa
Exposure time: 10.0 s

IHC: ICAD Antibody (Ab109205)

ICAD Antibody (Ab109205) - IHC
DAB staining on IHC
Samples: Human Breast cancer Tissue
Primary Ab: 10 µg/mL ICAD Antibody (Ab109205)
Second Ab: 2 µg/mL HRP-Linked Caprine Anti-Rabbit IgG Polyclonal Antibody.

IHC: ICAD Antibody (Ab109205)

ICAD Antibody (Ab109205) - IHC
DAB staining on IHC
Samples: Human Kidney Tissue
Primary Ab: 10 µg/mL ICAD Antibody (Ab109205)
Second Ab: 2 µg/mL HRP-Linked Caprine Anti-Rabbit IgG Polyclonal Antibody.

IF/ICC: ICAD Antibody (Ab109205)

ICAD Antibody (Ab109205) - IF/ICC

IF analysis of ICAD Antibody (Ab109205) in HeLa cells. The cells were fixed with 4% PFA for 10 minutes, permeabilized with 0.3% Triton X-100 (I743383) for 10 minutes and then blocked with 5% BSA for 1 hour at room temperature. Cells were stained with ICAD Antibody (Ab109205) at 1/200 dilution in blocking buffer overnight at 4°C, and then incubated with Goat Anti-Rabbit IgG H&L (AF488) (Ab181954) at a dilution of 1/1000 for 1 hour at room temperature in the dark (green). Cells were counterstained with DAPI (blue).

Applicazione

Applicazione

Dilution info

IHC

5-20 µg/mL

WB

0.01-2 µg/mL

IF/ICC

2.5 µg/mL

Obiettivi associati (umani)

DFFA Tbio DNA fragmentation factor subunit alpha (0 Activities)

Discover Target: DFFA

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

Application Protocols

Tested/validated applications, recommended dilutions, antigen retrieval, positive control tissues/cell lines, and detection systems: Not specified for this item; refer to CoA/Spec Sheet.

Note: Until item-specific protocols are provided, users should perform small-scale titrations and matrix checks to optimize conditions for their assay format.

Biological Roles

General background on ICAD (inhibitor of caspase-activated DNase; protein product commonly referred to as ICAD):

• Functional overview (literature):
  • ICAD forms a complex with CAD (caspase-activated DNase), acting as a chaperone and inhibitor. During apoptosis, caspase-3 (and related caspases) cleave ICAD, releasing active CAD to fragment chromosomal DNA into oligonucleosomal units.
  • ICAD exists as long (ICAD-L) and short (ICAD-S) isoforms with distinct regulatory properties.
• Cellular localization (literature):
  • Primarily cytosolic/nucleoplasmic; upon apoptotic signaling, processing enables CAD nuclear translocation for DNA fragmentation.
• Pathway context (literature):
  • Integral to the execution phase of apoptosis; intersects with caspase cascades and DNA damage responses.
• Research significance (general):
  • Monitoring ICAD levels/cleavage status serves as a biochemical readout of caspase activity and apoptotic progression in cell biology and biochemical assays.

Note: The above biological information is general context on the target protein and not a performance claim for this specific antibody. Species reactivity, epitope, and application suitability for SKU Ab109205 are not specified and should be confirmed experimentally or via the CoA/Spec Sheet.

Buffer Applications

This SKU is an antibody reagent; classic buffer formulation guidance is relevant for its handling and assays rather than for buffering per se.

General buffer guidance (not item-specific):

• Reconstitution/dilution:
  • Use sterile PBS (10–50 mM phosphate, 100–150 mM NaCl, pH 7.2–7.5) or TBS (20–50 mM Tris, 100–150 mM NaCl, pH 7.4–8.0), depending on downstream application. For NHS-ester conjugations, use amine-free buffers (phosphate, bicarbonate) during labeling.
• Additives (if compatible with your assay):
  • 0.01–0.05% Tween-20 can reduce nonspecific adsorption in washes.
  • 1–5% trehalose or 10–50% glycerol may improve freeze–thaw stability for storage at ≤−20 °C; verify compatibility with your assay.
  • Avoid azide if using peroxidase-based detection or live cells.
• Blocking/assay buffers:
  • For immunoassays, block with 1–5% non-interfering protein (BSA, casein) or commercial blockers; since this product is Carrier Free, blocking is added by the user as needed.

Item-specific buffer composition is not provided; consult the CoA/Spec Sheet for definitive recommendations and any restrictions.

Green Alternatives

• Waste minimization:
  • Use Carrier Free antibody to perform on-demand conjugations, reducing the need for multiple pre-conjugated formats that may expire unused.
  • Aliquot to extend shelf life and prevent repeated freeze–thaw waste.
• Buffer choices (greener practices, general):
  • Prefer phosphate- or HEPES-based buffers over volatile organic solvents; avoid azide if downstream live-cell work is intended to reduce toxic preservative waste.
• Packaging and cold-chain efficiency:
  • Consolidate orders to reduce shipments requiring ice packs/dry ice. Plan experiments to minimize reshipments and associated packaging waste.

Comparison (general, not item-specific):

• Carrier-containing formulations vs Carrier Free:
  • Carrier Free reduces animal-derived additives (e.g., BSA) and can simplify downstream purification after conjugation, lowering consumable use. Tradeoff: potentially reduced stability at very low concentrations—consider adding inert, non-animal stabilizers post-conjugation if compatible with your workflow.
• In-lab practices:
  • Employ reusable cold blocks and secondary containment; use low-adsorption plastics to reduce protein loss without resorting to excess stabilizers.

Pharmaceutical Uses

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

General formulation/manufacturing context for antibodies (not item-specific):

• As analytical reagents in bioprocess development and QC, antibodies may be used in ELISA or Western blot assays to monitor target protein expression in cell lines or process intermediates.
• In characterization workflows, Carrier Free antibodies can be custom-conjugated to reporters (e.g., HRP, biotin, fluorophores) for in-process analytics.
• No pharmacopeial monograph or excipient status is claimed for this item. Any GMP or clinical manufacturing use would require additional qualification and documentation not provided here.

Item-specific regulatory/quality claims beyond the listed grades (Carrier Free, ExactAb™, Validated, High Performance) are not specified for this SKU.

Physical Properties

• Appearance/form: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration: Not specified for this item; refer to CoA/Spec Sheet.
• Formulation/buffer: Carrier Free (item-specific grade); absence of carrier proteins such as BSA is implied by the grade. Other excipients (salts, preservatives) are not specified for this item.
• pH/osmolality: Not specified for this item; refer to CoA/Spec Sheet.
• Extinction coefficient: Not specified for this item; refer to CoA/Spec Sheet.
• Protein content determination: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general values (context, not item-specific):

• Typical IgG molecular mass ~150 kDa; Fab fragments ~50 kDa; scFv/VHH formats can be ~12–30 kDa depending on design.
• Typical antibody isoelectric points (pI) range ~6–9 depending on isotype and sequence.
• Storage temperatures commonly −20 °C to −80 °C for long term; working aliquots often kept at 2–8 °C for short term to minimize freeze–thaw cycling.

Solubility (general): antibodies are water-soluble proteins; solubility and stability depend strongly on buffer composition (pH 6.5–7.5, 50–150 mM salts) and presence/absence of stabilizers. Specific solubility for this item is not provided.

Quality and Grades

Item-specific grades provided: Carrier Free; ExactAb™; Validated; High Performance.

• Carrier Free (item-specific):
  • Indicates the antibody is supplied without carrier proteins such as BSA, gelatin, or casein. Benefits include compatibility with downstream conjugations (enzymes, fluorophores), labeling chemistries, mass spectrometry, and functional assays where carriers interfere.
  • Practical implications: Lower background in certain applications; users should add their own stabilizers if needed. Adsorptive losses can be higher at low µg/mL—pre-block plastics or add inert protein after conjugation if compatible with your use.
• ExactAb™ (brand/quality notation):
  • Denotes a high-consistency antibody preparation and/or well-characterized manufacturing control (general description). Refer to CoA/Spec Sheet for the exact definition used for this SKU.
• Validated (item-specific descriptor):
  • Asserts that performance was evaluated against defined criteria. The specific test methods, applications, and species validated are not specified for this item; consult CoA/Spec Sheet for details.
• High Performance (item-specific descriptor):
  • Indicates elevated purity or activity relative to standard research-grade antibodies. Concrete metrics (purity %, residual aggregates, endotoxin, host contaminants) are not specified for this item.

Quality documentation:

• For lot-specific information (purity, residual preservatives, concentration, isotype, clone, endotoxin), see the Certificate of Analysis/Specification Sheet. Do not infer unstated attributes.

Reaction and Applications

No manufacturer application methods are provided for this item. The following outlines general laboratory uses of antibodies targeting ICAD for researchers planning experiments.

• Immunodetection modalities (general guidance):
  • Western blotting to detect ICAD polypeptides under reducing/non-reducing conditions; antigen retrieval is not applicable to WB but denaturation and proper transfer conditions are critical due to molecular size of ICAD isoforms.
  • Immunoprecipitation or co-immunoprecipitation to enrich ICAD and associated complexes; optimize lysis buffers to preserve protein–protein interactions.
  • Immunofluorescence/ICC to localize ICAD in cells; fixation (paraformaldehyde) and permeabilization (e.g., saponin or Triton X-100) require titration to preserve epitope accessibility.
  • ELISA or bead-based assays to quantify ICAD in lysates; selection of capture/detection pairs depends on epitope non-overlap.
• Conjugation/use in multimodal assays:
  • Carrier Free format is compatible with enzymatic (HRP/AP), fluorescent (organic dyes), or biotin conjugation. Remove unreacted label by desalting or size exclusion chromatography prior to use.
• Controls and validation (general best practice):
  • Include positive controls with known ICAD expression and negative/knockdown controls where feasible.
  • Perform titration curves to identify the antibody’s working range; verify linearity for quantitative readouts.

Note: The above are general applications for anti-ICAD antibodies and are not a guarantee of performance for this specific SKU. Consult the CoA/Spec Sheet for item-validated applications and conditions.

Reaction Conditions

There are no synthetic reaction conditions for this biological reagent. The following are general, literature-based handling conditions for antibody–antigen binding in common assays; they are not item-specific performance guarantees.

• Incubation (general ranges):
  • Western blot: primary antibody 0.05–2 µg/mL in blocking buffer; 1 h at room temperature or overnight at 4 °C with gentle agitation.
  • Immunocytochemistry/IF: 0.5–10 µg/mL; 1 h at room temperature or overnight at 4 °C; optimize fixation/permeabilization for epitope accessibility.
  • Immunoprecipitation: 1–10 µg antibody per mg lysate protein; 1–4 h at 4 °C; use Protein A/G beads compatible with the antibody isotype (isotype for this item is not specified).
  • ELISA: coating 0.5–5 µg/mL capture antibody; detection antibody titrated to achieve desired dynamic range.
• Buffers/conditions:
  • Neutral pH (7.2–7.6), isotonic saline; include detergents (0.05% Tween-20) for washes to reduce nonspecific binding.
  • Avoid harsh detergents or high denaturant concentrations unless the assay requires it (e.g., SDS-PAGE before WB transfer).
• Controls:
  • Include no-primary and isotype controls (if isotype known); use positive/negative biological controls for ICAD expression.

Please confirm any item-validated conditions in the CoA/Spec Sheet once available.

Safety and Handling

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to the SDS for authoritative safety information.
• Research Use: For research use only (as stated). Not for diagnostic, therapeutic, or in vivo use.
• General precautions (good laboratory practice):
  • Wear appropriate PPE (lab coat, safety glasses, disposable gloves).
  • Avoid ingestion, inhalation, and contact with skin/eyes. Handle as a potentially bioactive protein solution.
  • Work on ice for extended handling to preserve activity; avoid microbial contamination.
• Incompatibilities/conditions to avoid:
  • Avoid repeated freeze–thaw cycles (item-specific). Aliquot upon first thaw.
  • Avoid strong oxidants, extreme pH, and elevated temperatures that can denature proteins.
  • Do not vortex aggressively; gentle mixing helps maintain conformational integrity.
• First-aid overview (general):
  • Skin/eye contact: Rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
  • Inhalation: Move to fresh air; obtain medical advice if symptoms occur.
  • Ingestion: Rinse mouth; seek medical attention.
• Spill/cleanup: Absorb with inert material, clean with water/detergent; disinfect surfaces where appropriate. Dispose according to institutional and local regulations.
• SDS: Always consult the product-specific SDS for final guidance on hazards, disposal, and transport.

Solvent Selection

This product is an antibody reagent and is not used as an organic solvent. Classic solvent selection is not applicable.

Practical guidance (general, for handling):

• Aqueous buffers are used for dilution/reconstitution (e.g., PBS or Tris-buffered saline, pH ~7.2–7.5). Additives like 0.01–0.1% Tween-20 and 0.1–1% inert proteins or sugars may improve stability; choose only if compatible with your downstream assay.
• For conjugation workflows, Carrier Free format is advantageous; use amine-free buffers (e.g., phosphate or bicarbonate) for NHS ester labeling and avoid primary amine buffers (e.g., Tris) during the conjugation step.
• For live-cell applications, avoid preservatives (e.g., azide) and low-salt conditions; verify that the product formulation matches your needs (not specified for this item—confirm in CoA/Spec Sheet).

Storage and Reconstitution

• Item-specific storage conditions:
  • Store at −20 °C.
  • Avoid repeated freezing and thawing; aliquot upon first thaw to minimize activity loss.
  • Shipped in an ice chest with ice pads to maintain cold chain.
• Reconstitution (general guidance; item-specific instructions not provided):
  • If supplied lyophilized: reconstitute with sterile, antibody-grade water or buffer (e.g., PBS) to the desired concentration. Gently invert to mix; avoid vigorous vortexing.
  • If supplied in solution: keep on ice during handling; dilute in assay-compatible buffer containing appropriate blockers only if required by the application.
• Stabilization tips (general):
  • For long-term storage at −20 °C, consider adding cryoprotectants such as 10–50% glycerol or 1–5% trehalose if compatible with your assay and permissible for this product; confirm formulation compatibility first.
  • Use low-protein-binding tubes and pre-chilled equipment to reduce adsorptive losses.
• Freeze–thaw guidance:
  • Prepare single-use aliquots; avoid more than one freeze–thaw cycle where possible. Thaw on ice and keep cold during setup.

For definitive instructions on concentration, formulation, and reconstitution volume for SKU Ab109205, consult the lot-specific CoA/Spec Sheet.

Structure and Identity

ICAD Antibody (SKU: Ab109205) is an affinity reagent directed against the ICAD antigen for research use.

• Item-specific identifiers:
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • InChIKey: Not applicable to proteins/antibodies; not specified for this item.
  • SMILES: Not applicable to proteins/antibodies; not specified for this item.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Structural class (general, literature):
  • Antibody immunoglobulin (protein macromolecule). Typical IgG is a Y‑shaped heterotetramer composed of two heavy and two light chains linked by disulfides, with variable domains forming the antigen-binding paratope.
  • Functional regions include Fab domains (antigen binding) and Fc region (effector/interaction with Fc receptors) when present; exact isotype/format for this item is not specified.
• Epitope/clone/isotype: Not specified for this item; refer to CoA/Spec Sheet.

2D/primary structure description (general): immunoglobulins contain β‑sandwich immunoglobulin domains with complementarity-determining regions (CDRs) in the variable domains that confer antigen specificity. The precise sequence and CDR composition for this antibody are not provided in the product data.

Synthetic Utility

Not typically applicable. This product is a biological antibody reagent rather than a small-molecule or synthetic building block. For experimental relevance, see the sections on Reaction & Applications, Buffer Applications, and Application Protocols (once specified).

Target Specificity

• Antigen/Target name: ICAD (as indicated by product name)
• Clone, isotype, host species, species reactivity, epitope mapping, and cross-reactivity: Not specified for this item; refer to CoA/Spec Sheet.

No additional target specificity details are provided in the product data.