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Product Info Summary
| SKU: | PB9756 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Cdc25C Antibody Picoband®
SKU/Catalog Number
PB9756
PB0805 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cdc25C Antibody Picoband® catalog # PB9756. Tested in WB, ICC/IF, Flow Cytometry 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-Cdc25C Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9756)
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 Cdc25C recombinant protein (Position: K175-P473). Human Cdc25C shares 78.2% amino acid (aa) sequence identity with mouse Cdc25C.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
PB9756 is reactive to CDC25C in Human, Mouse, Rat
Observed Molecular Weight
53 kDa
Calculated molecular weight
53.4 kDa
Background of CDC25C
M-phase inducer phosphatase 3 is an enzyme that in humans is encoded by the CDC25C gene. This gene is highly conserved during evolution and it plays a key role in the regulation of cell division. The encoded protein is a tyrosine phosphatase and belongs to the Cdc25 phosphatase family. It directs dephosphorylation of cyclin B-bound CDC2 (CDK1) and triggers entry into mitosis. Also, it is thought to suppress p53-induced growth arrest. Multiple alternatively spliced transcript variants of this gene have been described, however, the full-length nature of many of them is not known.
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
PB9756 is guaranteed for Flow Cytometry, IF, 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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human 293T whole cell, human Hela whole cell, human HepG2 whole cell, human K562 whole cell
ICC/IF: U2OS cell
FCM: K562 cell
Validation Images & Assay Conditions
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Western blot analysis of Cdc25C using anti-Cdc25C antibody (PB9756).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 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 Hela whole cell lysates.
Lane 3: human HepG2 whole cell lysates.
Lane 4: human K562 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-Cdc25C antigen affinity purified polyclonal antibody (PB9756) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Cdc25C at approximately 53 kDa. The expected band size for Cdc25C is at 53 kDa.
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IF analysis of Cdc25C using anti-Cdc25C antibody (PB9756) and anti-Tubulin Alpha antibody (M03989-3).
Cdc25C was detected in immunocytochemical section of U2OS cell. 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-Cdc25C Antibody (PB9756) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of K562 cells using anti-Cdc25C antibody (PB9756).
Overlay histogram showing K562 cells stained with PB9756 (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-Cdc25C Antibody (PB9756, 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-Cdc25C Antibody Picoband® (PB9756)
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