Human/Mouse/Rat CDC25B Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat CDC25B. Four splice variants of CDC25B, with molecular weights of 61, 63, 65, and 67 kDa, are known. The immunogen selected for this antibody is common to all variants. Immunoreactivity consistent with at least 3 variants has been detected in Western blots. In Western blots, this antibody does not cross-react with recombinant human (rh) CDC25A or rhCDC25C.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human CDC25B
    Glu391-Gln580
    Accession # P30305
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.3 µg/mL
    See below
  • Immunocytochemistry
    10-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human/Mouse/Rat CDC25B by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, DA3 mouse myeloma cell line, and Nb2‑11 rat lymphoma cell line. PVDF membrane was probed with 0.3 µg/mL of Goat Anti-Human/Mouse/Rat CDC25B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1649) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CDC25B at approximately 61 - 67 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
CDC25B in HL‑60 Human Cell Line. CDC25B was detected in immersion fixed HL‑60 human acute promyelocytic leukemia cell line using Goat Anti-Human/Mouse/Rat CDC25B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1649) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CDC25B

Cell Division Cycle 25B (Cdc25B) phosphatase removes inorganic phosphate groups covalently attached to tyrosine, serine and threonine residues in proteins (1). Breast cancer patients bearing tumors containing high levels of Cdc25B have been found to have a greater incidence of aggressive, high-grade tumors than those with low Cdc25B levels (2). In cells, the levels of Cdc25B activity are highest during the G2/M transition of the cell cycle, where it is suspected to be involved in “checkpoint” control of cell cycle progression (3). Overexpression of Cdc25B reduces the G2/M cell cycle block caused by ionizing radiation (4). Although activated by phosphorylation, Ser323 phosphorylation causes the enzyme to bind the protein 14-3-3, preventing substrate access to the catalytic site (5). One of the major substrates of Cdc25B is Cdc2, a kinase that is activated by dephosphorylation (6). The recombinant protein is truncated to remove the N‑terminal regulatory domains and is fully active.

  • References:
    1. Draetta, G. and J. Eckstein (1997) Biochim. Biophys. Acta 1332:M53.
    2. Galaktionov, K. et al. (1995) Science 269:1575.
    3. Lammer, C. et al. (1998) J. Cell Sci. 111:2445.
    4. Miyata, H. et al. (2001) Cancer Res. 61:3188.
    5. Forrest, A. and B. Gabrielli (2001) Oncogene 20:4393.
    6. Gautier, J. et al. (1991) Cell 67:197.
  • Long Name:
    Cell Division Cycle 25B
  • Entrez Gene IDs:
    994 (Human)
  • Alternate Names:
    CDC25B; CDC25HU2; cell division cycle 25 homolog B (S. cerevisiae); cell division cycle 25 homolog B (S. pombe); cell division cycle 25B; Dual specificity phosphatase Cdc25B; EC 3.1.3.48; M-phase inducer phosphatase 2
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