DFFA/DFF45 Monoclonal antibody

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  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.

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  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 AffiniPure Goat Anti-Mouse IgG(H+L).

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  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 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.

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

Cat No. 68177-1-Ig

Clone No.1F6H9

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

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

Synonyms

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

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Tested Applications

• Positive WB detected in: LNCaP cells, HeLa cells, HEK-293 cells, HepG2 cells, Jurkat cells, K-562 cells
• Positive IF/ICC detected in: HEK-293 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/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:

  CatNo: Ag32657

  Product name: Recombinant human DFFA,DFF45 protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 9-328 aa of BC000037

  Sequence: GVPESGEIRTLKPCLLRRNYSREQHGVAASCLEDLRSKACDILAIDKSLTPVTLVLAEDGTIVDDDDYFLCLPSNTKFVALASNEKWAYNNSDGGTAWISQESFDVDETDSGAGLKWKNVARQLKEDLSSIILLSEEDLQMLVDAPCSDLAQELRQSCATVQRLQHTLQQVLDQREEVRQSKQLLQLYLQALEKEGSLLSKQEESKAAFGEEVDAVDTGISRETSSDVALASHILTALREKQAPELSLSSQDLELVTKEDPKALAVALNWDIKKTETVQEACERELALRLQQTQSLHSLRSISASKASPPGDLQNPKRARQ

• Full Name: DNA fragmentation factor, 45kDa, alpha polypeptide
• Calculated Molecular Weight: 37 kDa
• Observed Molecular Weight: 42 kDa
• GenBank Accession Number: BC000037
• Gene Symbol: DFFA
• Gene ID (NCBI): 1676
• RRID: AB_2935268
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: O00273
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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.