Anti-Cdc25B Antibody Picoband™

Product Name

Anti-Cdc25B Antibody Picoband™

View all CDC25B Antibodies

SKU/Catalog Number

PB9488

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Cdc25B Antibody Picoband™ catalog # PB9488. Tested in IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Cdc25B Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9488)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Cdc25B recombinant protein (Position: M119-L248). Human Cdc25B shares 71% and 68.2% amino acid (aa) sequence identity with mouse and rat Cdc25B, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9488 is reactive to CDC25B in Human, Mouse, Rat

Applications

PB9488 is guaranteed for IHC, WB Boster Guarantee

Observed Molecular Weight

65 kDa

Calculated molecular weight

64.987kDa

Background of CDC25B

Central to the onset of mitosis in all eukaryotic cells is the CDC2 protein kinase, the activity of which is negatively regulated by phosphorylation and positively activated by dephosphorylation. The latter function is carried out by a specific phosphatase, CDC25. At least 3 human CDC25 genes code for the A, B, and C forms of CDC25. CDC25B is mapped to 20p13. P38 kinase has a critical role in the initiation of a G2 delay after ultraviolet radiation. Inhibition of p38 blocks the rapid initiation of this checkpoint in both human and murine cells after ultraviolet radiation. In vitro, p38 binds and phosphorylates CDC25B at serines 309 and 361, and CDC25C at serine-216; phosphorylation of these residues is required for binding to 14-3-3 proteins. In vivo, inhibition of p38 prevents both phosphorylation of CDC25B at serine-309 and 14-3-3 binding after ultraviolet radiation, and mutation of this site is sufficient to inhibit the checkpoint initiation. Regulation of CDC25B phosphorylation by p38 is a critical event for initiating the G2/M checkpoint after ultraviolet radiation.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat, Human, By Heat
Western blot, 0.1-0.5μg/ml, Rat, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Cdc25B using anti-Cdc25B antibody (PB9488).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Rat Liver Tissue Lysate at 50ug,
Lane 2: Rat Testis Tissue Lysate at 50ug.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Cdc25B antigen affinity purified polyclonal antibody (Catalog # PB9488) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Cdc25B at approximately 65 kDa. The expected band size for Cdc25B is at 65 kDa.

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Figure 2. IHC analysis of Cdc25B using anti-Cdc25B antibody (PB9488).
Cdc25B was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Cdc25B Antibody (PB9488) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of Cdc25B using anti-Cdc25B antibody (PB9488).
Cdc25B was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Cdc25B Antibody (PB9488) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Gene/Protein Information For CDC25B (Source: Uniprot.org, NCBI)

Alternative 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 CDC25B cell division cycle 25B M-phase inducer phosphatase 2|CDC25 homolog B|dual specificity phosphatase Cdc25B

For more info on CDC25B, check out the CDC25B Infographic

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In this infographic, you will see the following information for CDC25B: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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