Anti-Cyclin A2/CCNA2 Antibody Picoband™

Product Name

Anti-Cyclin A2/CCNA2 Antibody Picoband™

View all Cyclin A2 Antibodies

SKU/Catalog Number

PB9424

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Cyclin A2/CCNA2 Antibody Picoband™ catalog # PB9424. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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 A2/CCNA2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9424)

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 A2 recombinant protein (Position: A10-K168). Human Cyclin A2 shares 74.5% amino acid (aa) sequence identity with mouse Cyclin A2.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9424 is reactive to CCNA2 in Human, Mouse, Rat

Applications

PB9424 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

55 kDa

Calculated molecular weight

48.551kDa

Background of Cyclin A2

Cyclin A2, known as CCNA2, is mapped to 4q27. 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. In contrast to cyclin A1, which is present only in germ cells, this cyclin is expressed in all tissues tested. This cyclin binds and activates CDC2 or CDK2 kinases, and thus promotes both cell cycle G1/S and G2/M transitions. And Cyclin A2 is synthesized at the onset of S phase and localizes to the nucleus, where the cyclin A2-CDK2 complex is implicated in the initiation and progression of DNA synthesis. Phosphorylation of CDC6 and MCM4 by the cyclin A2-CDK2 complex prevents re-replication of DNA during the cell cycle.

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, By Heat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Cyclin A2 using anti-Cyclin A2 antibody (PB9424).
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 K562 whole cell lysates,
Lane 2: human HL-60 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: rat stomach tissue lysates,
Lane 5: rat testis tissue lysates,
Lane 6: mouse stomach 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 A2 antigen affinity purified polyclonal antibody (Catalog # PB9424) 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 Cyclin A2 at approximately 55 kDa. The expected band size for Cyclin A2 is at 49 kDa.

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Figure 2. IHC analysis of Cyclin A2 using anti-Cyclin A2 antibody (PB9424).
Cyclin A2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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-Cyclin A2 Antibody (PB9424) 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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Figure 3. IF analysis of Cyclin A2 using anti-Cyclin A2 antibody (PB9424).
Cyclin A2 was detected in an 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 5 μg/mL rabbit anti-Cyclin A2 Antibody (PB9424) 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.

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Figure 4. Flow Cytometry analysis of U20S cells using anti-Cyclin A2 antibody (PB9424).
Overlay histogram showing U20S cells stained with PB9424 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cyclin A2 Antibody (PB9424, 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 (Red line) was also used as a control.

Gene/Protein Information For CCNA2 (Source: Uniprot.org, NCBI)

Alternative Names

CCN1CCNAcyclin-A; CCNA; CCNA2; Cyclin A2; Cyclin-A; cyclin-A2 CCNA2 CCN1, CCNA cyclin A2 cyclin-A2|cyclin-A

For more info on CCNA2, check out the CCNA2 Infographic

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In this infographic, you will see the following information for CCNA2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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