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PAPPA2 ELISA 试剂盒

PAPPA2 适用: 人 Colorimetric Sandwich ELISA 0.1-10 ng/mL Plasma, Serum
产品编号 ABIN3073452
发货至: 中国
  • 抗原 See all PAPPA2 ELISA试剂盒
    PAPPA2 (Pappalysin 2 (PAPPA2))
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    0.1-10 ng/mL
    最低检测浓度
    0.1 ng/mL
    应用范围
    ELISA
    原理
    The PAPP-A2 enzyme linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of PAPP-A2 in human serum and other biological fluids. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.
    样品类型
    Plasma, Serum
    Analytical Method
    Quantitative
    灵敏度
    0.071 ng/mL
    组件
    • PAPP-A2 Calibrator A/Sample Diluent
    • PAPP-A2 Calibrators B thru F (Lyophilized)
    • PAPP-A2 Controls I & II (Lyophilized)
    • Anti-PAPP-A2 Antibody Coated Microtitration Strips
    • PAPP-A2 Assay Buffer
    • PAPP-A2 Biotin Conjugate Diluent
    • PAPP-A2 Biotin Conjugate Concentrate
    • PAPP-A2 Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
    • TMB Chromogen Solution
    • Stopping Solution
    • Wash Concentrate A
    试剂未包括
    1. Microtitration plate reader capable of absorbance measurement at 450nm, 405nm and 630nm.
    2. Microtitration orbital plate shaker.
    3. Microtitration plate washer.
    4. Semi-automated/manual precision pipette to deliver 10-250 μL.
    5. Vortex mixer.
    6. Deionized water.
    7. Disposable 12 x 75 mm culture tubes.
    8. Tight fitting 12 x 75 mm tube racks.
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    Discover our top product PAPPA2 ELISA Kit
  • 样本量
    50 μL
    实验时间
    2.5 h
    板类型
    Pre-coated
    试剂准备
    1. PAPP-A2 Calibrators B-F and Controls I & II: Tap and reconstitute PAPP-A2 Calibrators B-F and Controls I & II each with 1 mL deionized water. Solubilize for 10 minutes, mix well and use after reconstitution.
    2. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature when stored in a tightly sealed bottle.
    3. PAPP-A2 Antibody-Biotin Conjugate Solution: The PAPP-A2 Antibody-Biotin Conjugate Concentrate should be diluted at a of 1 part conjugate to 50 parts of PAPP-A2 Conjugate Diluent, according ratio to the number of wells used. If an entire plate is to be used pipet exactly 220 μL of the Concentrate in to 11 mL of the PAPP-A2 Conjugate Diluent.
    4. Microtitration Wells: Select the number of coated wells required for the assay. The remaining unused wells should be placed in the resealable pouch with a desiccant. The pouch must be resealed to protect from moisture.
    样品收集
    • Serum is the recommended sample type.
    • Use the following recommendations for handling, processing and storing blood samples.
      • Allow samples to clot for two hours at room temperature or overnight at 4oC and follow blood collection tube manufacturer's recommendations for centrifugation. Keep tubes stoppered at all times. Within two hours after centrifugation, transfer at least 500 μL of cell free sample to a storage tube. Tightly stopper the tube immediately.
      • Samples if used within 24 hours may be stored at 4 °C, otherwise samples must be stored at -20 °C or -80 °C to avoid loss of bioactivity and contamination.
      • Remove residual fibrin and cellular matter prior to analysis.
      • Avoid assaying lipemic, hemolyzed or icteric samples
      • Each laboratory should determine the acceptability of its own blood collection tubes and serum separation products.
      • Avoid repeated freezing and thawing of samples. Thaw samples no more than 3 times.
    实验流程

    Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. NOTE: All serum samples reading higher than the highest calibrator should be thoroughly mixed and diluted in the 0 ng/mL PAPP-A2 Cal A/Sample diluent prior to assay. For pregnancy serum specimens: Dilute specimens 1:20 with the Cal A/Sample diluent before assay. Do not dilute Calibrators or Controls.

    1. Mark the microtitration strips to be used.
    2. Pipette 50 μL of the Calibrator, Controls and Unknowns to the appropriate wells.
    3. Add 50 μL of the PAPP-A2 Assay Buffer to each well using a repeater pipette.
    4. Incubate the wells, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 1 hour at room temperature (23 ± 2 °C).
    5. With 30-40 minutes remaining of incubation time, prepare the PAPP-A2 Antibody-Biotin Conjugate Solution by diluting the PAPP-A2 Biotin Conjugate Concentrate in PAPP-A2 Conjugate Diluent as described under the Preparation of the Reagents section of this insert.
    6. Aspirate and wash each well 5 times with Wash Solution (350 μL/per well) using an automatic microplate washer.
    7. Add 100 μL of the Antibody-Biotin Conjugate solution to each well using a repeater pipette.
    8. Incubate the wells, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 1 hour at room temperature (23 ± 2 °C).
    9. Aspirate and wash each well 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
    10. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
    11. Incubate the wells, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature (23 ± 2 °C).
    12. Aspirate and wash each well 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
    13. Add 100 μL of the TMB chromogen solution to each well using a repeater pipette. Avoid exposure to direct sunlight.

    结果分析

    NOTE: The results in this package insert were calculated by plotting the log optical density (OD) data on the y-axis and log PAPP-A2 concentration on X- axis using a cubic regression curve-fit. Alternatively, log vs. log quadratic regression curve-fit can be used. Other data reduction methods may give slightly different results.

    1. Optimum results can be obtained at incubation temperature of 23 ± 2 °C.
    2. Calculate the mean absorbance for each calibrator, Control, or Unknown. Plot the log of the mean absorbance readings for each of the Calibrators along the y-axis versus log of the PAPP-A2 concentrations in ng/mL along the x-axis, using a cubic regression curve-fit.
    3. Determine the PAPP-A2 concentrations of the Controls and unknowns from the calibration curve by matching their mean absorbance readings with the corresponding PAPP-A2 concentrations.
    4. Any sample reading higher than the higest Calibrator should be appropriately diluted with the 0 ng/mL (CAL A/Sample diluent) and re- assayed. For pregnancy samples: Dilute 1:20 with the 0 ng/mL (Cal A/Sample diluent) before assay. Multiply the value by a dilution factor.
    5. Any sample reading lower than the analytical sensitivity should be reported as such.

    实验精密度
    Reproducibility of the PAPP-A2 ELISA assay was determined using four serum pools for a total of 12 assays. At concentrations between 1.0 and 3.1ng/mL, the CV's ranged between 4.1 and 4.6%.
    限制
    仅限研究用
  • 注意事项
    For Research Use Only. Not for use in diagnostic procedures. The following precautions should be observed: a) Follow good laboratory practice. b) Use personal protective equipment. Wear lab coats and disposable gloves when handling immunoassay materials. c) Handle and dispose of all reagents and material in compliance with applicable regulations. WARNING: Potential Biohazardous Material This reagent may contain some human source material (e.g. serum) or materials used in conjunction with human source materials. Handle all reagents and patient samples at a Biosafety Level 2, as recommended for any potentially infectious human material in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 5th Edition, 20079. WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium azide10 as a preservative. Pro-Clean 400 and Sodium azide in concentrated amounts are irritants to skin and mucous membranes. For further information regarding hazardous substances in the kit, please refer to the MSDS.
    储存条件
    4 °C
  • 抗原 See all PAPPA2 ELISA试剂盒
    PAPPA2 (Pappalysin 2 (PAPPA2))
    别名
    PAPP-A2 (PAPPA2 产品)
    别名
    PAPP-A2 ELISA Kit, PLAC3 ELISA Kit, Pappe ELISA Kit, PAPP-E ELISA Kit, PAPPE ELISA Kit, si:dkey-39f8.1 ELISA Kit, pappalysin 2 ELISA Kit, Pappa2 ELISA Kit, PAPPA2 ELISA Kit, pappa2 ELISA Kit
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