DBF4 Recombinant Monoclonal Antibody

Code CSB-RA155202A0HU
Size US$210
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  • Western Blot
    Positive WB detected in: PC3 whole cell lysate, Hela whole cell lysate, 293T whole cell lysate, HepG2 whole cell lysate, U937 whole cell lysate
    All lanes: DBF4 antibody at 1:2000
    Secondary
    Goat polyclonal to rabbit IgG at 1/50000 dilution
    Predicted band size: 77, 27 kDa
    Observed band size: 72-95 kDa
  • Immunofluorescence staining of A549 cell with CSB-RA155202A0HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 551-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
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Product Details

Uniprot No.
Target Names
Alternative Names
Protein DBF4 homolog A (Activator of S phase kinase) (Chiffon homolog A) (DBF4-type zinc finger-containing protein 1), DBF4, ASK DBF4A ZDBF1
Species Reactivity
Human
Immunogen
A synthesized peptide derived from human DBF4
Immunogen Species
Homo sapiens (Human)
Conjugate
Non-conjugated
Clonality
Monoclonal
Isotype
Rabbit IgG
Clone No.
24D8
Purification Method
Affinity-chromatography
Concentration
It differs from different batches. Please contact us to confirm it.
Buffer
Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
Form
Liquid
Tested Applications
ELISA, WB, IF
Recommended Dilution
Application Recommended Dilution
WB 1:500-1:2000
IF 1:50-1:200
Troubleshooting and FAQs
Storage
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time
Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Description

The synthesis of the DBF4 recombinant monoclonal antibody involves a stringent and well-defined process to ensure its quality and specificity. Initially, B cells are isolated from an immunized animal, with the synthesized peptide derived from human DBF4 serving as the immunogen. Subsequently, total RNA is extracted from the isolated B cells and converted into cDNA through reverse transcription. The DBF4 antibody genes are amplified using PCR with specific primers targeting the antibody constant regions and then inserted into an expression vector. This vector is introduced into host cells, enabling the production of the DBF4 recombinant monoclonal antibody. The antibody is harvested from the cell culture supernatant and purified using affinity chromatography, resulting in a highly purified preparation. Rigorous characterization assays, including ELISA, WB, and IF analysis, are performed to validate the antibody's specificity and functionality, ensuring its precise binding to human DBF4 protein.

Usage
For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function
Regulatory subunit for CDC7 which activates its kinase activity thereby playing a central role in DNA replication and cell proliferation. Required for progression of S phase. The complex CDC7-DBF4A selectively phosphorylates MCM2 subunit at 'Ser-40' and 'Ser-53' and then is involved in regulating the initiation of DNA replication during cell cycle.
Gene References into Functions
  1. Both CDC7 and DBF4 promoters bind E2F, suggesting that increased E2F activity in RB1 mutant cancers promotes increased DDK expression. Surprisingly, increased DDK expression levels are also correlated with both increased chemoresistance and genome-wide mutation frequencies. PMID: 28448802
  2. Histone H3 lysine 9 (H3K9) acetylation was most prevalent when the Dbf4 transcription level was highest whereas the H3K9me3 level was greatest during and just after replication. PMID: 27341472
  3. we propose that phosphorylation of TOP2A by CDC7/DBF4 in early S-phase prevents its localization and/or activity at centromeres, and inhibition of TOP2A function could be relevant to prevent premature separation of centromeric DNA. PMID: 27407105
  4. The anti-invasive and cell cycle arrest-inducing effects of nitrogen-containing bisphosphonates are not DBF4 mediated in human breast cancer cells. PMID: 24287290
  5. Dbf4 is direct downstream target of ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and Rad3-related (ATR) protein to regulate intra-S-phase checkpoint. PMID: 22123827
  6. bipartite interaction between Cdc7 and Dbf4/ASK subunits facilitates ATP binding and substrate recognition by the Cdc7 kinase. PMID: 21536671
  7. Dbf4 regulates the Cdc5 Polo-like kinase through a distinct non-canonical binding interaction PMID: 21036905
  8. Data document the role of DBF4 as a key player in nitrogen-containing bisphosphonate-induced cytotoxicity, thus explaining the effects on the cell-cycle. PMID: 19744312
  9. The interaction with LEDGF relieves autoinhibition of Cdc7-ASK kinase, imposed by the C terminus of ASK. PMID: 19864417
  10. results suggest that E2F regulates the ASK promoter through an atypical mode of recognition of the target site PMID: 12015319
  11. The HuDbf4 gene has of 12 exons over a 33-kb region. The MluI cell-cycle box is needed for core promoter activation. The Sp1 upregulator & HES-1 repressor coordinately determine promoter efficiency. Transcription initiations occur at -220, -235 and -245. PMID: 12420215
  12. Results demonstrate a DNA damage checkpoint that targets Cdc7/Dbf4 protein kinase. PMID: 12535533
  13. Cdc7-Dbf4 kinase efficiently phosphorylates p150. PMID: 16826239
  14. Taken together, these results indicate that Cdc7/Dbf4 phosphorylation of MCM2 is essential for the initiation of DNA replication in mammalian cells. PMID: 16899510
  15. Overexpression of Dbf4 abrogates the S checkpoint response to ultraviolet radiation but not ionizing radiation. PMID: 17276990
  16. There is the differential regulation of a novel gene, namely ASK/Dbf4, in melanoma and suggest that up-regulation of ASK/Dbf4 is a novel molecular determinant with prognostic relevance that confers a proliferative advantage in cutaneous melanoma. PMID: 17768177
  17. Cdc7/Dbf4 kinase activity inhibition affects specific phosphorylation sites on MCM2 in cancer cells PMID: 18286467
  18. ASK/Dbf4, a novel cell survival gene in melanoma is transcriptionally regulated by E2F1. PMID: 18503552
  19. A strong origin of replication at the DBF4 promoter locus, which contains two initiation zones, two origin recognition complex (ORC) binding sites and two DNase I-hypersensitive regions, is identified. PMID: 18536724
  20. increased Cdc7-Dbf4 abundance may be a common occurrence in human malignancies PMID: 18714392
  21. These results indicate that Ddk functions as an upstream regulator to monitor S-phase checkpoint signaling. PMID: 19111665

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Subcellular Location
Nucleus.
Tissue Specificity
Highly expressed in testis and thymus. Expressed also in most cancer cells lines.
Database Links

HGNC: 17364

OMIM: 604281

KEGG: hsa:10926

STRING: 9606.ENSP00000265728

UniGene: Hs.485380

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