Mouse Phospho AKT S473 (p-AKT) ELISA Kit

SKU: orb1736551

特色

描述

The Mouse Phospho AKT S473 (p-AKT) ELISA Kit uses a sandwich enzyme immunoassay to quantify Phospho AKT S473 (p-AKT) in mouse tissue homogenates, cell lysates and other biological fluids. The kit has a detection range of 156.25-10000 pg/mL and a sensitivity of 49.6 pg/mL.

研究领域

Signal Transduction

图片与验证

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Item 1 of 1

应用说明

standard: 10000 pg/mL. Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mouse p-AKT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse p-AKT. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mouse p-AKT, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mouse p-AKT in the samples is then determined by comparing the OD of the samples to the standard curve

关键属性

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反应性	Mouse
样本类型	tissue homogenates, cell lysates and other biological fluids
检测类型	Sandwich
检测时间	3.5h
范围	156.25-10000 pg/mL
灵敏度	49.6 pg/mL
浓度	10000 pg/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	This procedure is for reference only. 1. After the kit equilibrates to room temperature, add standards or samples to each well and incubate. 2. Discard liquid, add wash buffer to each well, wash the plate three times, and blot dry on clean absorbent paper. 3. Add biotinylated antibody working solution to each well and incubate. 4. Discard liquid, add wash buffer to each well, wash the plate three times, and blot dry on clean absorbent paper. 5. Add streptavidin-HRP working solution to each well and incubate._x000b_6. Discard liquid, add wash buffer to each well, wash the plate five times, and blot dry on clean absorbent paper. 7. Add TMB substrate solution to each well and incubate in the dark. 8. 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	请参阅操作手册中的储存说明
有效期	6 months from date of receipt.
免责声明	For research use only

别名

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p-AKT

质量保证

Explore bioreagents carefree to elevate your research. All our products are rigorously tested for performance. If a product does not perform as described on its datasheet, our scientific support team will provide expert troubleshooting, a prompt replacement, or a refund. For full details, please see our 条款与条件和选购指南.

实验方案信息

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按应用筛选

ELISA (1)

按种属筛选

Mouse (1)

Fatemah A Alherz 1, Asmaa Saleh 1, Mona Y Alsheikh 2, Hany M Borg 3, Ahmed M Kabel 4, Maaly A Abd Elmaaboud Shikonin mitigates cyclophosphamide-induced cardiotoxicity in mice: the role of sirtuin-1, NLRP3 inflammasome, autophagy, and apoptosis J Pharm Pharmacol, (2024)

PubMed: 39245439

应用

ELISA

反应性

Mouse