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Human TOP2A / Topoisomerase II Alpha ELISA Kit

SKU:
HUFI02040
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P11388
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
TOP2A, TOP2, TP2A, DNA topoisomerase II, alpha isozyme, DNA topoisomerase, ATP-hydrolyzing, DNA topoisomerase 2-alpha, TOP2DNA gyrase, topoisomerase, DNA II alpha, 170kD, topoisomerase, DNA II alpha 170kDa
Reactivity:
Human
Research Area:
Cell Biology
€599
Frequently bought together:

Description

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Human TOP2A/Topoisomerase II Alpha ELISA Kit

The Human TOP2A (Topoisomerase II Alpha) ELISA Kit is specifically designed for the quantitative measurement of TOP2A levels in human samples, including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit provides accurate and reliable results for a variety of research applications.TOP2A is an important enzyme involved in DNA replication and repair, making it a key player in cell division and growth. Dysregulation of TOP2A has been linked to various diseases, including cancer and autoimmune disorders.

As a crucial biomarker, studying TOP2A levels can provide valuable insights into disease progression and potential treatment options.Whether you are researching cancer biology, drug development, or cell signaling pathways, the Human TOP2A ELISA Kit is an essential tool for identifying and quantifying TOP2A levels in human samples, driving innovative research and advancements in the field.

Product Name:Human TOP2A / Topoisomerase II Alpha ELISA Kit
Product Code:HUFI02040
Size:96 Assays
Alias:TOP2A, TOP2, TP2A, DNA topoisomerase II, alpha isozyme, DNA topoisomerase, ATP-hydrolyzing, DNA topoisomerase 2-alpha, TOP2DNA gyrase, topoisomerase, DNA II alpha, 170kD, topoisomerase, DNA II alpha 170kDa
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human TOP2A concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human TOP2A and the recovery rates were calculated by comparing the measured value to the expected amount of Human TOP2A in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-10297
EDTA plasma(n=5)92-9996
UFH plasma(n=5)85-10495
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human TOP2A and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)87-104%87-105%93-105%
EDTA plasma(n=5)93-98%82-94%87-100%
UFH plasma(n=5)95-99%89-99%83-99%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP11388
UniProt Protein Function:Control of topological states of DNA by transient breakage and subsequent rejoining of DNA strands. Topoisomerase II makes double-strand breaks. Essential during mitosis and meiosis for proper segregation of daughter chromosomes. May play a role in regulating the period length of ARNTL/BMAL1 transcriptional oscillation.
NCBI Summary:This gene encodes a DNA topoisomerase, an enzyme that controls and alters the topologic states of DNA during transcription. This nuclear enzyme is involved in processes such as chromosome condensation, chromatid separation, and the relief of torsional stress that occurs during DNA transcription and replication. It catalyzes the transient breaking and rejoining of two strands of duplex DNA which allows the strands to pass through one another, thus altering the topology of DNA. Two forms of this enzyme exist as likely products of a gene duplication event. The gene encoding this form, alpha, is localized to chromosome 17 and the beta gene is localized to chromosome 3. The gene encoding this enzyme functions as the target for several anticancer agents and a variety of mutations in this gene have been associated with the development of drug resistance. Reduced activity of this enzyme may also play a role in ataxia-telangiectasia. [provided by RefSeq, Jul 2010]
UniProt Code:P11388
NCBI GenInfo Identifier:13959709
NCBI Gene ID:7153
NCBI Accession:P11388.3
UniProt Secondary Accession:P11388,Q71UN1, Q71UQ5, Q9HB24, Q9HB25, Q9HB26, Q9UP44 Q9UQP9, B2RTS1,
UniProt Related Accession:P11388
Molecular Weight:182,681 Da
NCBI Full Name:DNA topoisomerase 2-alpha
NCBI Synonym Full Names:topoisomerase (DNA) II alpha
NCBI Official Symbol:TOP2A
NCBI Official Synonym Symbols:TOP2; TP2A
NCBI Protein Information:DNA topoisomerase 2-alpha
UniProt Protein Name:DNA topoisomerase 2-alpha
UniProt Synonym Protein Names:DNA topoisomerase II, alpha isozyme
Protein Family:DNA topoisomerase
UniProt Gene Name:TOP2A
UniProt Entry Name:TOP2A_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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