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Product Info Summary
| SKU: | A00953 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Topoisomerase II alpha/TOP2A Antibody Picoband®
SKU/Catalog Number
A00953
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Topoisomerase II alpha/TOP2A Antibody Picoband® catalog # A00953. Tested in Flow Cytometry, IF, IHC, 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-Topoisomerase II alpha/TOP2A Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00953)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human Topoisomerase II alpha/TOP2A, identical to the related mouse and rat sequences.
*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
A00953 is reactive to TOP2A in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
174 kDa
Calculated molecular weight
50408 MW
Background of TOP2A
The human topoisomerase II enzyme is encoded by a single-copy gene which is mapped to 17q21-q22. The TOP2A gene spans approximately 30 kb and contains 35 exons. Furthermore, DNA topoisomerases are enzymes that control and alter the topologic states of DNA in both prokaryotes and eukaryotes. Topoisomerase II from eukaryotic cells catalyzes the relaxation of supercoiled DNA molecules, catenation, decatenation, knotting, and unknotting of circular DNA. It appears likely that the reaction catalyzed by topoisomerase II involves the crossing-over of 2 DNA segments. There are about 100,000 molecules of topoisomerase II per HeLa cell nucleus, constituting about 0.1% of the nuclear extract1. DNA topoisomerase II-alpha is associated with the pol II holoenzyme and is a required component of chromatin-dependent coactivation. Specific inhibitors of topoisomerase II blocked transcription on chromatin templates, but did not affect transcription on naked templates. Addition of purified topoisomerase II-alpha reconstituted chromatin-dependent activation activity in reactions with core pol II2.
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
A00953 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, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human A549 whole cell, human U2OS whole cell, human HEK293 whole cell, human K562 whole cell
IHC: human Lung cancer tissue, human rectal cancer tissue
ICC/IF: U20S cell
FCM: 293T cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of TOP2A using anti-TOP2A antibody (A00953).
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: human A549 whole cell lysates,
Lane 2: human U2OS whole cell lysates,
Lane 3: human HEK293 whole cell lysates,
Lane 4: human K562 whole cell lysates.
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-TOP2A antigen affinity purified polyclonal antibody (Catalog # A00953) 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 TOP2A at approximately 174KD. The expected band size for TOP2A is at 174KD.
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Figure 2. IHC analysis of TOP2A using anti-TOP2A antibody (A00953).
TOP2A was detected in paraffin-embedded section of human Lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TOP2A Antibody (A00953) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IHC analysis of TOP2A using anti-TOP2A antibody (A00953).
TOP2A was detected in paraffin-embedded section of human rectal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TOP2A Antibody (A00953) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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Figure 4. Flow Cytometry analysis of 293T cells using anti-TOP2A antibody (A00953).
Overlay histogram showing 293T cells stained with A00953 (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-TOP2A Antibody (A00953,1μg/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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Figure 5. IF analysis of TOP2A using anti-TOP2A antibody (A00953).
TOP2A was detected in immunocytochemical section of U20S 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-TOP2A Antibody (A00953) 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.
Protein Target Info & Infographic
Gene/Protein Information For TOP2A (Source: Uniprot.org, NCBI)
Gene Name
TOP2A
Full Name
DNA topoisomerase 2-alpha
Weight
50408 MW
Superfamily
type II topoisomerase family
Alternative Names
DNA topoisomerase 2-alpha; DNA topoisomerase II, alpha isozyme; TOP2A; TOP2 TOP2A TOP2, TP2A DNA topoisomerase II alpha DNA topoisomerase 2-alpha|DNA gyrase|DNA topoisomerase (ATP-hydrolyzing)|DNA topoisomerase II, 170 kD|DNA topoisomerase II, alpha isozyme|topoisomerase (DNA) II alpha 170kDa
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on TOP2A, check out the TOP2A Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for TOP2A: 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].
Specific Publications For Anti-Topoisomerase II alpha/TOP2A Antibody Picoband® (A00953)
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Customer Q&As
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5 Customer Q&As for Anti-Topoisomerase II alpha/TOP2A Antibody Picoband®
Question
We ordered your anti-Topoisomerase II alpha/TOP2A antibody for Flow Cytometry on leukemic t-cell a few years ago. I am using human, and I plan to use the antibody for WB next. My lab would like examining leukemic t-cell as well as cervix carcinoma erythroleukemia in our next experiment. Could you please give me some suggestion on which antibody would work the best for WB?
Verified Customer
Verified customer
Asked: 2020-03-18
Answer
I looked at the website and datasheets of our anti-Topoisomerase II alpha/TOP2A antibody and I see that A00953 has been tested on human in both Flow Cytometry and WB. Thus A00953 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2020-03-18
Question
My boss were well pleased with the WB result of your anti-Topoisomerase II alpha/TOP2A antibody. However we have observed positive staining in ovarian carcinoma cytoplasm. using this antibody. Is that expected? Could you tell me where is TOP2A supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-06-24
Answer
From literature, ovarian carcinoma does express TOP2A. Generally TOP2A expresses in cytoplasm. Regarding which tissues have TOP2A expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 10942766, 16964243, 17081983, 17924679, 18220336, 18669648, 18691976, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Boster Scientific Support
Answered: 2019-06-24
Question
We are currently using anti-Topoisomerase II alpha/TOP2A antibody A00953 for human tissue, and we are well pleased with the IHC-P results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on horse tissues as well?
Verified Customer
Verified customer
Asked: 2018-10-29
Answer
The anti-Topoisomerase II alpha/TOP2A antibody (A00953) has not been validated for cross reactivity specifically with horse 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 horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-10-29
Question
I am looking for using your anti-Topoisomerase II alpha/TOP2A antibody for cellular response to dna damage stimulus studies. Has this antibody been tested with western blotting on a549 whole cell lysates? We would like to see some validation images before ordering.
B. Johnson
Verified customer
Asked: 2017-04-06
Answer
Thank you for your inquiry. This A00953 anti-Topoisomerase II alpha/TOP2A antibody is validated on human a549, a549 whole cell lysates, u2os whole cell lysates, k562 whole cell lysates, 293t cells. It is guaranteed to work for Flow Cytometry, IHC-P, 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: 2017-04-06
Question
We have observed staining in human cervix carcinoma erythroleukemia. Any tips? Is anti-Topoisomerase II alpha/TOP2A antibody supposed to stain cervix carcinoma erythroleukemia positively?
J. Carter
Verified customer
Asked: 2014-01-27
Answer
Based on literature cervix carcinoma erythroleukemia does express TOP2A. Based on Uniprot.org, TOP2A is expressed in heart, ovarian carcinoma, cervix carcinoma, leukemic t-cell, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have TOP2A expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 10942766, 16964243, 17081983, 17924679, 18220336, 18669648, 18691976, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Boster Scientific Support
Answered: 2014-01-27
