Anti-CDC25B Antibody Picoband™

Product Name

Anti-CDC25B Antibody Picoband™

View all CDC25B Antibodies

SKU/Catalog Number

A01899-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CDC25B Antibody Picoband™ catalog # A01899-1. Tested in Flow Cytometry, ELISA, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-CDC25B Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01899-1)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human CDC25B recombinant protein (Position: M1-H486).

*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

A01899-1 is reactive to CDC25B in Human, Mouse, Rat

Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Observed Molecular Weight

70 kDa

Calculated molecular weight

40895 MW

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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Applications

A01899-1 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

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.

Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Positive Control

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of CDC25B using anti-CDC25B antibody (A01899-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: human MOLT-4 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human Hela whole cell lysates,
Lane 6: human Hacat whole cell lysates.
Lane 7: human SiHa whole cell lysates.
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 # A01899-1) 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 70 kDa. The expected band size for CDC25B is at 65 kDa.

Click image to see more details

Figure 2. Western blot analysis of CDC25B using anti-CDC25B antibody (A01899-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat testis tissue lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse testis tissue lysates.
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 # A01899-1) 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 70 kDa. The expected band size for CDC25B is at 65 kDa.

Click image to see more details

Figure 3. Flow Cytometry analysis of U87 cells using anti-CDC25B antibody (A01899-1).
Overlay histogram showing U87 cells stained with A01899-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CDC25B Antibody (A01899-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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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