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Product Info Summary
| SKU: | PB9485 |
|---|---|
| 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-Cyclin A1/CCNA1 Antibody Picoband®
SKU/Catalog Number
PB9485
PB0515 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cyclin A1/CCNA1 Antibody Picoband® catalog # PB9485. Tested in Flow Cytometry, IF, 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-Cyclin A1/CCNA1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9485)
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 Cyclin A1 recombinant protein (Position: D64-E263). Human Cyclin A1 shares 78.5% and 79.5% amino acid (aa) sequence identity with mouse and rat Cyclin A1, respectively.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9485 is reactive to CCNA1 in Human, Mouse, Rat
Observed Molecular Weight
52 kDa
Calculated molecular weight
52.4 kDa
Background of CCNA1
Cyclin-A1 is a protein that in humans is encoded by the CCNA1 gene. The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. The cyclin encoded by this gene was shown to be expressed in testis and brain, as well as in several leukemic cell lines, and is thought to primarily function in the control of the germline meiotic cell cycle. This cyclin binds both CDK2 and CDC2 kinases, which give two distinct kinase activities, one appearing in S phase, the other in G2, and thus regulate separate functions in cell cycle. Also this cyclin was found to bind to important cell cycle regulators, such as Rb family proteins, transcription factor E2F-1, and the p21 family proteins. Multiple transcript variants encoding different isoforms have been found for this gene.
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
PB9485 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, 5μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human 293T whole cell, human MCF-7 whole cell, human MCF-7 whole cell, human Hela whole cellrat kidney tissue, rat testis tissue, mouse kidney tissue, mouse testis tissue
ICC/IF: A549 cell
FCM: PC-3 cell
Validation Images & Assay Conditions
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Western blot analysis of Cyclin A1/CCNA1 using anti-Cyclin A1/CCNA1 antibody (PB9485).
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 MCF-7 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: rat testis tissue lysates,
Lane 6: mouse kidney tissue lysates,
Lane 7: 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-Cyclin A1/CCNA1 antigen affinity purified polyclonal antibody (Catalog # PB9485) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Cyclin A1/CCNA1 at approximately 52 kDa. The expected band size for Cyclin A1/CCNA1 is at 52 kDa.
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IF analysis of Cyclin A1/CCNA1 using anti-Cyclin A1/CCNA1 antibody (PB9485) and anti-Tubulin Alpha antibody (M03989-3).
Cyclin A1/CCNA1 was detected in immunocytochemical section of A549 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-Cyclin A1/CCNA1 Antibody (PB9485) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and FITC Conjugated Goat Anti-Mouse IgG (BA1101) 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 PC-3 cells using anti-Cyclin A1/CCNA1 antibody (PB9485).
Overlay histogram showing PC-3 cells stained with PB9485 (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-Cyclin A1/CCNA1 Antibody (PB9485, 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-Cyclin A1/CCNA1 Antibody Picoband® (PB9485)
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1 Customer Q&As for Anti-Cyclin A1/CCNA1 Antibody Picoband®
Question
We are currently using anti-Cyclin A1/CCNA1 antibody PB9485 for human tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human. Is it likely that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2017-08-21
Answer
The anti-Cyclin A1/CCNA1 antibody (PB9485) has not been tested for cross reactivity specifically with canine 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 canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-08-21
