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PLG ELISA 试剂盒

PLG 适用: 大鼠 Colorimetric Sandwich ELISA 0.391-400 ng/mL Cell Culture Cells, Plasma, Serum, Urine
产品编号 ABIN5564562
发货至: 中国
  • 抗原 See all PLG ELISA试剂盒
    PLG (Plasminogen (PLG))
    适用
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    大鼠
    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    0.391-400 ng/mL
    最低检测浓度
    0.391 ng/mL
    应用范围
    ELISA
    原理
    The AssayMax™ Rat Plasminogen ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of rat plasminogen in plasma, serum, urine, and cell culture samples. This assay employs a quantitative sandwich enzyme immunoassay technique that measures rat plasminogen in less than 4 hours. A polyclonal antibody specific for rat plasminogen has been pre-coated onto a 96-well microplate with removable strips. Plasminogen in standards and samples is sandwiched by the immobilized antibody and the biotinylated polyclonal antibody specific for rat plasminogen, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
    品牌
    AssayMax™
    样品类型
    Cell Culture Cells, Plasma, Serum, Urine
    Analytical Method
    Quantitative
    组件
    Rat Plasminogen Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against rat plasminogen. Sealing Tapes: Each kit contains 3 precut, pressure sensitive sealing tapes that can be cut to fit the format of the individual assay. Rat Plasminogen Standard: Rat plasminogen in a buffered protein base (800 ng, lyophilized). Biotinylated Rat Plasminogen Antibody (70x): A 70-fold concentrated biotinylated polyclonal antibody against rat plasminogen (105 l). EIA Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80 l). Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).
    试剂未包括
    Microplate reader capable of measuring absorbance at 405 nm. Pipettes (1-20 µL, 20-200 µL, and multiple channel). Deionized or distilled reagent grade water Incubator (37 °C)
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  • 板类型
    Pre-coated
    实验流程
    • Step 1. Add 50 μL of Standard or Sample per well. Incubate 2 hours.
    • Step 2. Wash, then add 50 μL of Biotinylated Antibody per well. Incubate 1 hour.
    • Step 3. Wash, then add 50 μL of SP Conjugate per well. Incubate 30 minutes.
    • Step 4. Wash, then add 50 μL of Chromogen Substrate per well. Incubate 12 minutes.
    • Step 5. Add 50 μL of Stop Solution per well. Read at 450 nm immediately.
    试剂准备

    Freshly dilute all reagents and bring all reagents to room temperature before use. EIA Diluent Concentrate (10x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. Dilute the EIA Diluent Concentrate 1:10 with reagent grade water. Store for up to 30 days at 2-8 °C.

    样品收集
    Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 3000 x g for 10 minutes. Dilute samples 1:20000 with EIA Diluent or within the range of 1:5000 - 1:50000. The undiluted samples can be stored at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles (EDTA or Heparin can also be used as an anticoagulant). Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 3000 x g for 10 minutes. Dilute samples 1:20000 with EIA Diluent or within the range of 1:5000 - 1:50000. The undiluted samples can be stored at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Cell Culture Supernatants: Collect cell culture media and centrifuge at 3000 x g for 10 minutes to remove debris. The user should determine the optimal dilution factor. Dilute cell culture media into EIA Diluent and assay. Store samples at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Urine: Collect urine using sample pot. Centrifuge samples at 800 x g for 10 minutes. Dilute urine 1:4 into EIA Diluent and assay. The undiluted samples can be stored at -20 °C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Refer to Sample Dilution Guidelines below for further instruction. Guidelines for Dilutions of 1:100 or Greater (for reference only, please follow the insert for specific dilution suggested) 1:100 1:10000 A) 4 μL sample: 396 μL buffer(100x) = 100 fold dilution Assuming the needed volume is less than or equal to 400 μL. A) 4 μL sample : 396 μL buffer (100x) B) 4 μL of A : 396 μL buffer (100x) = 10000 fold dilution Assuming the needed volume is less than or equal to 400 μL. 1:1000 1:100000 A) 4 μL sample : 396 μL buffer (100x) B) 24 μL of A : 216 μL buffer (10x) = 1000 fold dilution Assuming the needed volume is less than or equal to 240 μL. A) 4 μL sample : 396 μL buffer (100x) B) 4 μL of A : 396 μL buffer (100x) C) 24 μL of B : 216 μL buffer (10x) = 100000 fold dilution Assuming the needed volume is less than or equal to 240 μL.
    实验流程

    Prepare all reagents, standard solutions, and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20-25 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccants inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. 7 Add 50 l of Rat Plasminogen Standard or sample per well. Cover wells with a sealing tape and incubate for 2 hours. Start the timer after the last addition. Wash five times with 200 l of wash buffer manually. Invert the plate each time and decant the contents, hit 4-5 times on absorbent material to completely remove the liquid. If using a machine, wash six times with 300 l of wash buffer and then invert the plate, decanting the contents, hit 4-5 times on absorbent material to completely remove the liquid. Add 50 l of Biotinylated Rat Plasminogen antibody to each well and incubate for 1 hour. Wash the microplate as described above. Add 50 l of Streptavidin-Peroxidase Conjugate per well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash the microplate as described above. Add 50 l of Chromogen Substrate per well and incubate for 12 minutes or till the optimal blue color density develops. Gently tap the plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 l of stop solution to each well. The color will change from blue to yellow. Read the absorbance on a microplate reader at a wavelength of 450 nm immediately. If wavelength correction is available, subtract readings at 570 nm from those at 450 nm to correct optical imperfections. Otherwise, read the plate at 450 nm only. Please note that some unstable black particles may be generated at high concentration points after stopping the reaction for about 10 minutes, which will reduce the readings.

    结果分析
    • Calculate the mean value of the duplicate or triplicate readings for each standard and sample.
    • To generate a standard curve, plot the graph using the standard concentrations on the x-axis and the corresponding mean 450 nm absorbance (OD) on the y-axis. The best-fit line can be determined by regression analysis using log-log or four-parameter logistic curve-fit.
    • Determine the unknown sample concentration from the standard curve and multiply the value by the dilution factor.
    限制
    仅限研究用
  • 注意事项
    This product is for Research Use Only and is Not For Use In Diagnostic Procedures. Prepare all reagents (working diluent buffer, wash buffer, standard, biotinylated antibody, and SP conjugate) as instructed, prior to running the assay. Prepare all samples prior to running the assay. The dilution factors for the samples are suggested in this insert. However, the user should determine the optimal dilution factor. Spin down the SP conjugate vial and the biotinylated antibody vial before opening and using contents. Rat Plasminogen ELISA Kit Catalog No. ERP1200-1 Sample insert for reference use only The Stop Solution is an acidic solution. The kit should not be used beyond the expiration date.
    储存条件
    4 °C,-20 °C
    储存方法
    Upon arrival, immediately store components of the kit at recommended temperatures up to the expiration date. Store SP conjugate and Biotinylated Antibody at -20°C. Store Microplate, Diluent Concentrate (10x), Wash Buffer, Stop Solution, and Chromogen Substrate at 2-8°C. Unused microplate wells may be returned to the foil pouch with the desiccant packs and resealed. May be stored for up to 30 days in a vacuum desiccator. Diluent (1x) may be stored for up to 30 days at 2-8°C. Store Standard at 2-8°C before reconstituting with Diluent and at -20°C after reconstituting with Diluent.
  • 抗原 See all PLG ELISA试剂盒
    PLG (Plasminogen (PLG))
    别名
    Plasminogen (PLG 产品)
    别名
    wu:fb70e09 ELISA Kit, PLG ELISA Kit, LPA ELISA Kit, plg ELISA Kit, Ab1-346 ELISA Kit, AI649309 ELISA Kit, Pg ELISA Kit, plasminogen ELISA Kit, plg ELISA Kit, PLG ELISA Kit, Plg ELISA Kit
    背景
    Plasminogen is a single chain glycoprotein zymogen that is synthesized in the liver and circulated in plasma with a molecular weight of 90 kDa. The N- terminal portion of the molecule is made up of five kringle domains that bind to fibrin. The native molecule has an amino-terminal glutamic acid, known as glu-plasminogen, but this can undergo proteolytic cleavage by plasmin to lys- plasminogen (1). The inactive proenzyme plasminogen is converted to the active enzyme plasmin that ultimately digests fibrin. Tissue-type plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA) catalyzes the activation of plasminogen, while plasminogen activator inhibitors (PAIs) inhibit the activation (2-3).
    基因ID
    85253
    UniProt
    Q01177
    途径
    Complement System, Lipid Metabolism
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