Human HIF1A (Hypoxia-inducible factor 1-alpha) Quick ELISA Kit

SKU: orb2568477

描述

图片与验证

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Item 1 of 1

关键属性

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反应性	Human
样本类型	Serum, plasma, cell culture supernatant, cell lysate or tissue lysate, other biological fluid samples
检测类型	Sandwich
检测时间	120 minutes
范围	0.156-10ng/ml
灵敏度	0.094ng/ml

步骤与表现

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Assay Principle	The kit is based on a sandwich enzyme immunoassay principle. The microtiter plate is pre-coated with a capture antibody specific to the target analyte. Standards or samples are added to the wells, followed by a biotin-conjugated detection antibody specific for the analyte. Avidin conjugated to horseradish peroxidase (HRP) is then added and incubated. After addition of the TMB substrate, color develops only in wells containing the analyte bound to the detection antibody and HRP–avidin complex. The reaction is stopped with an acidic solution, and absorbance is measured at 450 nm ± 10 nm. The analyte concentration in the samples is determined by comparison with a standard curve.
Kit Components	1. ELISA Microplate 2. Standards 3. Detection Antibody 4. HRP-Streptavidin Conjugate 5. TMB Substrate 6. Dilution buffers 7. Stop Solution 8. Wash Buffer 9. Plate Sealers 10. Manual
Reagent Preparation	1. Wash Buffer: Prepare the 1X Wash Buffer using distilled water according to the manual. 2. Standard: Perform gradient dilution according to the instructions in the manual. 3. Other Concentrated Reagents: Dilute the concentrated reagents using the Dilution Buffers provided in the kit to 1 X working solutions as instructed in the manual. Always use a clean pipette tip for each different solution.
Assay Procedure	After the kit equilibrates to room temperature, add standards or samples to each well and incubate. Discard liquid, add wash buffer to each well, wash the plate twice, and blot dry on clean absorbent paper. Add biotinylated antibody working solution to each well and incubate. Discard liquid, add wash buffer to each well, wash the plate three times, and blot dry on clean absorbent paper. Add streptavidin-HRP working solution to each well and incubate. Discard liquid, add wash buffer to each well, wash the plate five times, and blot dry on clean absorbent paper. Add TMB substrate solution to each well and incubate in the dark. Add stop solution to each well, mix thoroughly, and immediately read OD at 450 nm.
Materials Required	1. Microplate readers 2. Centrifuge 3. Incubator 4. Automated plate washer 5. Single-channel or multi-channel high-precision pipettes 6. Disposable pipette tips 7. Sterile tubes 8. Eppendorf tubes 9. Absorbent paper 10. Loading slots
Precision	Intra-assay Precision (Precision within an assay): CV% < 8% Intra-assay precision was evaluated by testing multiple replicates of samples within the same plate. Inter-assay Precision (precision between assays): CV% < 10% Inter-assay precision was evaluated by testing samples across different plates.
Calculation of Results	1. Average the duplicate readings for each Standard, Control, and Sample, and subtract the mean optical density of the zero Standard. 2. Construct a standard curve by plotting the target concentration on the y-axis against absorbance on the x-axis and draw a curve through the data points. 3. Determine the sample concentration by substituting the OD450 value into the standard curve. For diluted samples, multiply the calculated value by the corresponding dilution factor.
Curve Fitting Softwares	1. Curve Expert 2. Thermo SkanIt RE 3. SciDAVis 4. LabPlot 5. ……

储存与操作

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储存	2 - 8 °C for 12 months
有效期	Please enquire.
免责声明	For research use only

别名

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质量保证

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常用数据库链接

基因符号

HIF1A

UniProt

Q16665

UniProt 详情

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暂无 UniProt 数据

实验方案信息

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Human HIF1A (Hypoxia-inducible factor 1-alpha) Quick ELISA Kit

描述

图片与验证

关键属性

步骤与表现

储存与操作

别名

质量保证

常用数据库链接

UniProt 详情

实验方案信息

可选规格