| 原理: |
This assay employs a two-site sandwich ELISA to quantitate NAT10 in samples. An antibody specific for NAT10 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any NAT10 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NAT10 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of NAT10 bound in the initial step. The color development is stopped and the intensity of the color is measured.
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| 产品概述: |
Cotransfection of HeLa cells with a reporter construct driven by the TERT promoter and ALP cDNA resulted in increased reporter activity. Cotransfection of the reporter with antisense ALP cDNA resulted decreased reporter activity. Cells overexpressing ALP showed higher telomerase activity, whereas cells with antisense ALP showed decreased telomerase activity. In vitro analysis showed that ALP specifically catalyzed acetylation of histones. The deduced 834-amino acid protein has a calculated molecular mass of 94 kD. ALP contains an N-acetyltransferase domain accompanied by an ATP/GTP-binding motif and an ATPase domain. Western blot analysis of HeLa cells detected ALP at an apparent molecular mass of 94 kD. Immunocytochemical analysis detected ALP mainly in HeLa cell nuclei.
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| 组份: |
Reagents |
Quantity |
Reagents |
Quantity |
Assay plate (96 Wells) |
1 |
Instruction manual |
1 |
Standard (lyophilized) |
2 |
Sample Diluent |
1 x 20 mL |
Biotin-Conjugate (concentrate 100 x) |
1 x 120 μL |
Biotin-Conjugate Diluent |
1 x 20 mL |
Streptavidin-HRP (concentrate 100 x) |
1 x 120 μL |
Streptavidin-HRP Diluent |
1 x 20 mL |
Wash Buffer (concentrate 25 x) |
1 x 20 mL |
Substrate Solution |
1 x 12 mL |
Stop Solution |
1 x 10 mL |
Adhesive Films |
4 |
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| 特异性: |
This assay has high sensitivity and excellent specificity for detection of Rat NAT10. No significant cross-reactivity or interference between Rat NAT10 and analogues was observed.
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| 回收率: |
Matrices listed below were spiked with certain level of recombinant Rat NAT10 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat NAT10 in samples.
Sample Type |
Number |
Recovery range (%) |
Average(%) |
Serum |
10 |
90-101 |
96 |
EDTA plasma |
10 |
89-97 |
93 |
Heparin plasma |
10 |
91-99 |
95 |
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| 精密度: |
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
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| 线性: |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat NAT10 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample Type |
1：2 |
1：4 |
1：8 |
1：16 |
Serum |
78-89% |
81-99% |
92-103% |
95-105% |
EDTA plasma |
91-101% |
90-98% |
93-101% |
91-98% |
Heparin plasma |
92-103% |
93-102% |
92-99% |
91-101% |
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| 稳定性: |
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
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| 样本收集及储存: |
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
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| 注意事项: |
Store at 2-8°C. Please refer to Instruction Manual.
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