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DFFA,DFF45 Monoclonal antibody

DFFA,DFF45 Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

1F6H9

Cat no : 68177-1-Ig

Synonyms

DFF45, DFFA, DFF45/DFFA, DFF-45, DFF1

Validation Data Gallery

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  • Western Blot (WB) analysis of various lysates using DFFA,DFF45 Monoclonal antibody (68177-1-Ig)

    WB analysis using 68177-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 68177-1-Ig (DFFA,DFF45 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

  • Immunofluorescence (IF) / fluorescent staining of HEK-293 cells using DFFA,DFF45 Monoclonal antibody (68177-1-Ig)

    IF Staining of HEK-293 using 68177-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using DFFA,DFF45 antibody (68177-1-Ig, Clone: 1F6H9 ) at dilution of 1:2000 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

  • Flow cytometry (FC) experiment of HEK-293 cells using DFFA,DFF45 Monoclonal antibody (68177-1-Ig)

    FC experiment of HEK-293 using 68177-1-Ig

    1x10^6 HEK-293 cells were intracellularly stained with 0.8 ug DFFA,DFF45 Monoclonal antibody (68177-1-Ig, Clone:1F6H9) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1)(red), or 0.8 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3) (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.

Tested Applications

Positive WB detected inLNCaP cells, HeLa cells, HEK-293 cells, HepG2 cells, Jurkat cells, K-562 cells
Positive IF/ICC detected inHEK-293 cells
Positive FC (Intra) detected inHEK-293 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunofluorescence (IF)/ICCIF/ICC : 1:1000-1:4000
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

68177-1-Ig targets DFFA,DFF45 in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity human, mouse, rat
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Immunogen DFFA,DFF45 fusion protein Ag32657
Full Name DNA fragmentation factor, 45kDa, alpha polypeptide
Calculated Molecular Weight 37 kDa
Observed Molecular Weight42 kDa
GenBank Accession NumberBC000037
Gene Symbol DFFA
Gene ID (NCBI) 1676
Conjugate Unconjugated
Form Liquid
Purification MethodProtein G purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Apoptosis is accompanied by shrinkage and fragmentation of the cells and nuclei and degradation of the chromosomal DNA into nucleosomal units. DNA fragmentation factor (DFF), heterodimer of 40-kDa (DFFB) and 45-kDa (DFFA) subunits, is one of the major endonucleases responsible for internucleosomal DNA cleavage during apoptosis. DFFA is the substrate for caspase-3 and triggers DNA fragmentation during apoptosis. DFF activated after cleavage of DFFA by caspase-3 followed by its dissociation from DFFB. DFFB has been found to trigger both DNA fragmentation and chromatin condensation during apoptosis. DFFA appears to be a 45-kDa protein despite the predicated MW of 37 kDa.

Protocols

Product Specific Protocols
WB protocol for DFFA,DFF45 antibody 68177-1-IgDownload protocol
IF protocol for DFFA,DFF45 antibody 68177-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols