FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of A431 cells using anti-DFFA/ICAD antibody (AAA125699).Overlay histogram showing A431 cells stained with AAA125699 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DFFA/ICAD Antibody (AAA125699, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.)
Rabbit anti-Human, Rat DFFA/ICAD Polyclonal Antibody | anti-DFFA antibody
Anti-DFFA/ICAD Antibody
Gene Names
DFFA; DFF1; ICAD; DFF-45
Reactivity
Human, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Western Blot
Purity
Immunogen affinity purified.
Synonyms
DFFA/ICAD, Antibody; Anti-DFFA/ICAD Antibody; DFFA; DFF1; DFF45; H13; DNA fragmentation factor subunit alpha; DNA fragmentation factor 45 kDa subunit; DFF-45; Inhibitor of CAD; ICAD; anti-DFFA antibody
Host
Rabbit
Reactivity
Human, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for DFFA/ICAD detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.005mg NaN3.
Applicable Applications for anti-DFFA antibody
ELISA (Direct ELISA), FCM/FACS (Flow Cytometry), WB (Western Blot)
Immunogen
E Coli-derived human DFFA/ICAD recombinant protein (Position: E14-S308).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.
FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of A431 cells using anti-DFFA/ICAD antibody (AAA125699).Overlay histogram showing A431 cells stained with AAA125699 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DFFA/ICAD Antibody (AAA125699, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.)
FCM/FACS (Flow Cytometry)
(Figure 2. Flow Cytometry analysis of A431 cells using anti-DFFA/ICAD antibody (AAA125699).Overlay histogram showing A431 cells stained with AAA125699 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DFFA/ICAD Antibody (AAA125699, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.)
WB (Western Blot)
(Figure 1. Western blot analysis of DFFA/ICAD using anti-DFFA/ICAD antibody (AAA125699).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 lysatesLane 2: human HEK293 whole cell lysatesLane 3: human HEPG2 whole cell lysatesLane 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 # AAA125699) at 0. 5 μg/mL overnight at 4 degree 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 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.)
WB (Western Blot)
(Figure 1. Western blot analysis of DFFA/ICAD using anti-DFFA/ICAD antibody (AAA125699).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 lysatesLane 2: human HEK293 whole cell lysatesLane 3: human HEPG2 whole cell lysatesLane 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 # AAA125699) at 0. 5 μg/mL overnight at 4 degree 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 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.)
Related Product Information for anti-DFFA antibody
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.
References
1. Abel, F., Sjoberg, R. -M., Ejeskar, K., Krona, C., Martinsson, T. Analyses of apoptotic regulators CASP9 and DFFA at 1p36. 2, reveal rare allele variants in human neuroblastoma tumours. Brit. J. Cancer 86: 596-604, 2002.
2. Inohara, N., Koseki, T., Chen, S., Wu, X., Nunez, G. CIDE, a novel family of cell death activators with homology to the 45 kDa subunit of the DNA fragmentation factor. EMBO J. 17: 2526-2533, 1998.
3. Leek, J. P., Carr, I. M., Bell, S. M., Markham, A. F., Lench, N. J. Assignment of the DNA fragmentation factor gene (DFFA) to human chromosome bands 1p36. 3-p36. 2 by in situ hybridization. Cytogenet. Cell Genet. 79: 212-213, 1997.
2. Inohara, N., Koseki, T., Chen, S., Wu, X., Nunez, G. CIDE, a novel family of cell death activators with homology to the 45 kDa subunit of the DNA fragmentation factor. EMBO J. 17: 2526-2533, 1998.
3. Leek, J. P., Carr, I. M., Bell, S. M., Markham, A. F., Lench, N. J. Assignment of the DNA fragmentation factor gene (DFFA) to human chromosome bands 1p36. 3-p36. 2 by in situ hybridization. Cytogenet. Cell Genet. 79: 212-213, 1997.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
36,522 Da
NCBI Official Full Name
DNA fragmentation factor subunit alpha isoform 1
NCBI Official Synonym Full Names
DNA fragmentation factor, 45kDa, alpha polypeptide
NCBI Official Symbol
DFFA
NCBI Official Synonym Symbols
DFF1; ICAD; DFF-45
NCBI Protein Information
DNA fragmentation factor subunit alpha; DFF45; inhibitor of CAD; DNA fragmentation factor 45 kDa subunit
UniProt Protein Name
DNA fragmentation factor subunit alpha
UniProt Gene Name
DFFA
UniProt Synonym Gene Names
DFF1; DFF45; DFF-45; ICAD
UniProt Entry Name
DFFA_HUMAN
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Product Notes
The DFFA dffa (Catalog #AAA125699) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-DFFA/ICAD Antibody reacts with Human, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's DFFA/ICAD can be used in a range of immunoassay formats including, but not limited to, ELISA (Direct ELISA), FCM/FACS (Flow Cytometry), WB (Western Blot). Researchers should empirically determine the suitability of the DFFA dffa for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "DFFA/ICAD, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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