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Product Info Summary
| SKU: | PB9104 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Cyclin B1/CCNB1 Antibody Picoband®
SKU/Catalog Number
PB9104
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cyclin B1/CCNB1 Antibody Picoband® catalog # PB9104. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. 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-Cyclin B1/CCNB1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9104)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Cyclin B1 recombinant protein (Position: M1-V433). Human Cyclin B1 shares 86% and 85% amino acid (aa) sequences identity with mouse and rat Cyclin B1, respectively.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9104 is reactive to CCNB1 in Human
Observed Molecular Weight
56 kDa
Calculated molecular weight
48.3 kDa
Background of CCNB1
CCNB also known as Cyclin B1, is a protein that in humans is encoded by the CCNB1 gene, and it is mapped to 5q13.2. The protein encoded by this gene is a regulatory protein involved in mitosis. The gene product complexes with p34 (cdc2) to form the maturation-promoting factor (MPF). Two alternative transcripts have been found, a constitutively expressed transcript and a cell cycle-regulated transcript, that is expressed predominantly during G2/M phase. The different transcripts result from the use of alternate transcription initiation sites. CCNB contributes to the switch-like all or none behavior of the cell in deciding to commit to mitosis. Its activation is well-regulated, and positive feedback loops ensure that once the cyclin B1-Cdk1 complex is activated, it is not deactivated.
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
PB9104 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
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: HELA whole cell, 293T whole cell, MCF-7 whole cell, COLO320 whole cell
ICC/IF: A431 cell
FCM: THP-1 cell
Validation Images & Assay Conditions
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Western blot analysis of CCNB1 using anti-CCNB1 antibody (PB9104).
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 50ug of sample under reducing conditions.
lane 1: HELA whole cell lysate,
lane 2: 293T whole cell lysate,
lane 3: MCF-7 whole cell lysate,
lane 4: COLO320 whole cell lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CCNB1 antigen affinity purified polyclonal antibody (Catalog # PB9104) 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 1:10000 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 CCNB1 at approximately 56KD.
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Flow Cytometry analysis of THP-1 cells using anti-CCNB1 antibody (PB9104).
Overlay histogram showing THP-1 cells stained with PB9104 (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-CCNB1 Antibody (PB9104,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of CCNB1 using anti-CCNB1 antibody (PB9104).
CCNB1 was detected in immunocytochemical section of A431 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 2μg/mL rabbit anti-CCNB1 Antibody (PB9104) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.
Specific Publications For Anti-Cyclin B1/CCNB1 Antibody Picoband® (PB9104)
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1 Customer Q&As for Anti-Cyclin B1/CCNB1 Antibody Picoband®
Question
I am interested in using your anti-Cyclin B1/CCNB1 antibody for mitotic cell cycle phase transition studies. Has this antibody been tested with western blotting on colo320 whole cell lysate? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-11-13
Answer
We appreciate your inquiry. This PB9104 anti-Cyclin B1/CCNB1 antibody is tested on hela whole cell lysate, 293t whole cell lysate, colo320 whole cell lysate. It is guaranteed to work for Flow Cytometry, IHC, ICC, WB in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-11-13
