Anti-DFFA/ICAD Antibody Picoband™

Product Name

Anti-DFFA/ICAD Antibody Picoband™

View all DFF45/ICAD Antibodies

SKU/Catalog Number

A03671-4

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-DFFA/ICAD Antibody Picoband™ catalog # A03671-4. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-DFFA/ICAD Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03671-4)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.005mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human DFFA/ICAD recombinant protein (Position: E14-S308).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A03671-4 is reactive to DFFA in Human, Rat

Applications

A03671-4 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

45 kDa

Calculated molecular weight

36.522kDa

Background of DFF45/ICAD

DNA fragmentation factor subunit alpha (DFFA), also known as Inhibitor of caspase-activated DNase (ICAD), is a protein that in humans is encoded by the DFFA gene. Apoptosis is a cell death process that removes toxic and/or useless cells during mammalian development. The apoptotic process is accompanied by shrinkage and fragmentation of the cells and nuclei and degradation of the chromosomal DNA into nucleosomal units. DNA fragmentation factor (DFF) is a heterodimeric protein of 40-kD (DFFB) and 45-kD (DFFA) subunits. DFFA is the substrate for caspase-3 and triggers DNA fragmentation during apoptosis. DFF becomes activated when DFFA is cleaved by caspase-3. The cleaved fragments of DFFA dissociate from DFFB, the active component of DFF. DFFB has been found to trigger both DNA fragmentation and chromatin condensation during apoptosis. Two alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of DFFA/ICAD using anti-DFFA/ICAD antibody (A03671-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HELA whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human HEPG2 whole cell lysates,
Lane 4: rat stomach tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DFFA/ICAD antigen affinity purified polyclonal antibody (Catalog # A03671-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DFFA/ICAD at approximately 45KD. The expected band size for DFFA/ICAD is at 45KD.

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Figure 2. Flow Cytometry analysis of A431 cells using anti-DFFA/ICAD antibody (A03671-4).
Overlay histogram showing A431 cells stained with A03671-4 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DFFA/ICAD Antibody (A03671-4, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For DFFA (Source: Uniprot.org, NCBI)

Alternative Names

DFF1; DFF1DNA fragmentation factor 45 kDa subunit; DFF45; DFF45DNA fragmentation factor subunit alpha; DFF-45DNA fragmentation factor, 45 kD, alpha polypeptide; DFFA; DNA fragmentation factor, 45kDa, alpha polypeptide; ICAD; Inhibitor of CAD DFFA DFF-45, DFF1, ICAD DNA fragmentation factor subunit alpha DNA fragmentation factor subunit alpha|DFF45|DNA fragmentation factor 45 kDa subunit|DNA fragmentation factor, 45kDa, alpha polypeptide|inhibitor of CAD

For more info on DFFA, check out the DFFA Infographic

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In this infographic, you will see the following information for DFFA: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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