Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CDC25B Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CDC25B
Glu391-Gln580
Accession # P30305

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat CDC25B Antibody

Detection of Human/Mouse/Rat CDC25B antibody by Western Blot.

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.

CDC25B antibody in HL-60 Human Cell Line by Immunocytochemistry (ICC).

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.

Applications for Human/Mouse/Rat CDC25B Antibody

Application
Recommended Usage

Immunocytochemistry

10-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells and HL‑60 human acute promyelocytic leukemia cell line

Western Blot

0.3 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, DA3 mouse myeloma cell line, and Nb2-11 rat lymphoma cell line

Reviewed Applications

Read 1 review rated 5 using AF1649 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

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

Entrez Gene IDs

994 (Human)

Gene Symbol

CDC25B

UniProt

P30305

Additional CDC25B Products

Product Documents for Human/Mouse/Rat CDC25B Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat CDC25B Antibody

For research use only

Related Research Areas

Cell Cycle
Phosphatases and Regulators

Citations for Human/Mouse/Rat CDC25B Antibody

Customer Reviews for Human/Mouse/Rat CDC25B Antibody (1)

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  • Human/Mouse/Rat CDC25B Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Colon cancer tissue
    Species: Human
    Verified Customer | Posted 09/22/2021
    Human/Mouse/Rat CDC25B Antibody AF1649

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Protocols

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