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Product Info Summary
| SKU: | A01899-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-CDC25B Antibody Picoband®
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01899-1 is reactive to CDC25B in Human, Mouse, Rat
Observed Molecular Weight
70 kDa
Calculated molecular weight
65.0 kDa
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.
Application & Images
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
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human 293T whole cell, human CACO-2 whole cell, human MOLT-4 whole cell, human Raji whole cell, human Hela whole cell, human Hacat whole cellhuman SiHa whole cell, rat liver tissue, rat brain tissue, rat kidney tissue, rat testis tissue, mouse liver tissue, mouse brain tissue, mouse kidney tissue, mouse testis tissue
FCM: U87 cell
Validation Images & Assay Conditions
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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
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
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.
Specific Publications For Anti-CDC25B Antibody Picoband® (A01899-1)
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