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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 (same clone as 68177-1-PBS)

    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. This data was developed using the same antibody clone with 68177-1-PBS in a different storage buffer formulation.

  • 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 (same clone as 68177-1-PBS)

    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 AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 68177-1-PBS in a different storage buffer formulation.

  • 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 (same clone as 68177-1-PBS)

    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 AffiniPure 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. This data was developed using the same antibody clone with 68177-1-PBS in a different storage buffer formulation.

  • Cytometric bead array standard curve of MP50354-1

    Cytometric bead array standard curve of MP50354-1

    Cytometric bead array standard curve of MP50354-1, DFFA,DFF45 Monoclonal Matched Antibody Pair, PBS Only. Capture antibody: 68177-1-PBS. Detection antibody: 68177-2-PBS. Standard:Ag32657. Range: 3.125-200 ng/mL.

  • Cytometric bead array standard curve of MP50354-2

    Cytometric bead array standard curve of MP50354-2

    Cytometric bead array standard curve of MP50354-2, DFFA,DFF45 Monoclonal Matched Antibody Pair, PBS Only. Capture antibody: 68177-3-PBS. Detection antibody: 68177-1-PBS. Standard:Ag32657. Range: 6.25-200 ng/mL.

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DFFA/DFF45 Monoclonal antibody, PBS Only (Capture/Detector)

DFFA/DFF45 Monoclonal Antibody for WB, IF/ICC, FC (Intra), Cytometric bead array, Indirect ELISA
Cat No. 68177-1-PBS
Clone No.1F6H9

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), Cytometric bead array, Indirect ELISA

Synonyms

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

Formulation:  PBS Only
PBS and Azide
PBS Only
Conjugate:  Unconjugated
Size/Concentration: 

-/ -

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Tested Applications

Recommended dilution

ApplicationDilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

68177-1-PBS targets DFFA/DFF45 as part of a matched antibody pair:

MP50354-1: 68177-1-PBS capture and 68177-2-PBS detection (validated in Cytometric bead array)

MP50354-2: 68177-3-PBS capture and 68177-1-PBS detection (validated in Cytometric bead array)

Unconjugated mouse monoclonal antibody pair in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

Tested Reactivity human, mouse, rat
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Immunogen DFFA/DFF45 fusion protein Ag32657 相同性解析による交差性が予測される生物種
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
UNIPROT IDO00273
Storage Buffer PBS only , pH 7.3
Storage ConditionsStore at -80°C.

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.