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Product Info Summary
| SKU: | PB9488 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Cdc25B Antibody Picoband®
SKU/Catalog Number
PB9488
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cdc25B Antibody Picoband® catalog # PB9488. Tested in Flow Cytometry, IF, IHC, ICC, 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
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 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
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.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9488 is reactive to CDC25B in Human, Mouse, Rat
Observed Molecular Weight
65 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
PB9488 is guaranteed for Flow Cytometry, IF, IHC, ICC, 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Jurkat whole cell lysates, human Raji whole cell lysates, human Hela whole cell lysates, human HepG2 whole cell lysates, rat liver tissue lysates, mouse liver tissue lysates
IHC: human stomach cancer tissue
IF: U2OS cell
FCM: HepG2 cell
Validation Images & Assay Conditions
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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. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse liver 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 # 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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IHC analysis of Cdc25B using anti-Cdc25B antibody (PB9488).
Cdc25B was detected in a paraffin-embedded section of human stomach cancer 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 2 μg/ml rabbit anti-Cdc25B Antibody (PB9488) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of Cdc25B using anti-Cdc25B antibody (PB9488).
Cdc25B was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-Cdc25B Antibody (PB9488) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of HepG2 cells using anti-Cdc25B antibody (PB9488).
Overlay histogram showing HepG2 cells stained with PB9488 (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 (PB9488, 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® (PB9488)
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4 Customer Q&As for Anti-Cdc25B Antibody Picoband®
Question
Will anti-Cdc25B antibody PB9488 work on monkey IHC with right hemisphere of cerebellum?
Verified Customer
Verified customer
Asked: 2020-04-02
Answer
Our lab technicians have not tested anti-Cdc25B antibody PB9488 on monkey. You can run a BLAST between monkey and the immunogen sequence of anti-Cdc25B antibody PB9488 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated monkey samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in monkey right hemisphere of cerebellum in IHC, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-04-02
Question
We are currently using anti-Cdc25B antibody PB9488 for human tissue, and we are well pleased with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2019-05-27
Answer
The anti-Cdc25B antibody (PB9488) has not been validated for cross reactivity specifically with zebrafish tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-05-27
Question
Would PB9488 anti-Cdc25B antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2017-06-14
Answer
You can see on the product datasheet, PB9488 anti-Cdc25B antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2017-06-14
Question
I was wanting to use your anti-Cdc25B antibody for WB for human cervix carcinoma on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human cervix carcinoma identification?
M. Krishna
Verified customer
Asked: 2013-07-31
Answer
As indicated on the product datasheet, PB9488 anti-Cdc25B antibody has been tested for IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human cervix carcinoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2013-07-31
