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Androgen Receptor (AR) ELISA Kit
概述
| 抗原: | Androgen receptor (AR) |
| 应用范围: | ELISA |
| 目标物种: | Rat (Rattus) |
| 规格: | |
| 价格: | 6.000,20 ¥ (加上运费和增值税, Invoice in EUR) |
| 货号: | ABIN416542 |
| 发货时间: | Ships within 10 to 15 Business days |
详细信息 » Androgen Receptor (AR) ELISA Kit
| 抗原 | Androgen receptor (AR) |
| 目标物种 | Rat (Rattus) |
| 特异性 | This assay has high sensitivity and excellent specificity for detection of rat AR. No significant cross-reactivity or interference was observed. |
| Sensitivity | The minimum detectable dose of rat AR is typically less than 0.06 ng/mL. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D. Value of 20 replicates of the zero standard added by their three standard deviations. Detection Range: 0.156-10 ng/mL. The standard curve concentrations used for the ELISAs were 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL. |
使用细节 » Androgen Receptor (AR) ELISA Kit
| Principle | The microtiter plate provided in this kit has been pre-coated with an antibody specific to AR. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for AR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain AR, biotin- conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm 2 nm. The concentration of AR in the samples is then determined by comparing the O.D. of the samples to the standard curve. |
| Protocol | SAMPLE COLLECTION & STORAGE: Other 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 for later use. Avoid repeated freeze/thaw cycles. Note: 1. Samples to be used within 5 days may be stored at 2-8°C , otherwise samples must stored at -20°C (1 month) or -80°C (2 months) to avoid loss of bioactivity and contamination. 2. When performing the assay slowly bring samples to room temperature. REAGENT PREPARATION: 1. Bring all kit components and samples to room temperature (18-25°C) before use. 2. Standard -- Reconstitute the Standard with 1.0 ml of Standard Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 10 ng/mL. Please prepare 7 tubes containing 0.5ml Standard Diluent and produce a double dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL. 3. Assay Diluent A and Assay Diluent B -- Dilute 6mL of Assay Diluent A or B Concentrate (2 ) with 6ml of deionized or distilled water to prepare 12 mL of Assay Diluent A or B. The prepared working dilution may not be frozen. 4. Detection Reagent A and Detection Reagent B -- Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or Assay Diluent B respectively(1:100). 5. Wash Solution - Dilute 20mL of Wash Solution Concentrate (30 ) with 580ml of deionized or distilled water to prepare 600 mL of Wash Solution (1 ). 6. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again. Note:1. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 directly. 2. Making serial dilution in the wells directly is not permitted. 3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μl for once pipetting. 4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used used used used only only only only once once once once. 5. If crystals have formed in the Wash Solution concentrate (30 ), warm to room temperature and mix gently until the crystals have completely dissolved. CALCULATION OF RESULTS: Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the AR concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation, such as curve expert 13.0. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. |
| 应用范围 | ELISA |
| Components | Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard (freeze dried) 2 Standard Diluent 1 × 20ml Detection Reagent A (green) 1 × 120μl Detection Reagent B (red) 1 × 120μl Assay Diluent A (2 x concentrate) 1 × 6ml Assay Diluent B (2 x concentrate) 1 × 6ml TMB Substrate 1 × 9ml Stop Solution 1 × 6ml Wash Buffer(30 x concentrate) 1 × 20ml |
| Material not included | 1. Microplate reader with 450 ± 10 nm filter. 2. Precision single and multi-channel pipettes and pipette tips with disposable tips. 3. Eppendorf Tubes for diluting samples. 4. Deionized or distilled water. 5. Absorbent paper for blotting the microtiter plate. 6. Container for Wash Solution |
| 研究领域 | Hormones, Receptors |
| 限制 | For Research Use only |
抗原
