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Albumin ELISA 试剂盒

ALB 适用: 小鼠 Colorimetric Competition ELISA Plasma
产品编号 ABIN612732
发货至: 中国
  • 抗原 See all Albumin (ALB) ELISA试剂盒
    Albumin (ALB)
    适用
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    小鼠
    检测方法
    Colorimetric
    实验类型
    Competition ELISA
    最低检测浓度
    300 ng/mL
    应用范围
    ELISA
    原理
    The AssayMax Mouse Albumin ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of mouse albumin in plasma and serum
    品牌
    AssayMax
    样品类型
    Plasma
    Analytical Method
    Quantitative
    组件
    Mouse Albumin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against mouse albumin. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes that can be cut to fit the format of the individual assay. Mouse Albumin Standard: Mouse albumin in a buffered protein base (150 µg, lyophilized). Biotinylated Albumin: 1 vial, lyophilized. MIx Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (90µl). 1 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 450 nm. Pipettes (1-20 µL, 20-200 µL, 200-1000µLand multiple channel). Deionized or distilled reagent grade water.
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  • 样本量
    25 μL
    实验时间
    < 2 h
    板类型
    Pre-coated
    实验流程
    This assay employs a quantitative competitive enzyme immunoassay technique that measures mouse albumin in less than 2 hours. A polyclonal antibody specific for mouse albumin has been pre-coated onto a 96-well microplate with removable strips. Albumin in standards and samples is competed by a biotinylated albumin sandwiched by the immobilized antibody and streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
    试剂准备

    Freshly dilute all reagents and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. MIx Diluent Concentrate (10x): Dilute the MIx Diluent 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. Standard Curve: Reconstitute the 150 g of albumin standard with 1.5 ml of MIx Diluent to generate a stock solution of 100 g/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare triplicate standard points by serially diluting the stock solution (100 g/ml) 1:4 with MIx Diluent to generate 25, 6.25, 1.563, 0.391 and 0.098 g/ml standard solutions. MIx Diluent serves as the zero standard (0 g/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [mAlbumin] ( g/ml) 1 part Stock (100 g/ml) + 3 parts MIx Diluent P1 25.000 P2 1 part P1 + 3 parts MIx Diluent 6.250 P3 1 part P2 + 3 parts MIx Diluent 1.563 P4 1 part P3 + 3 parts MIx Diluent 0.391 P5 1 part P4 + 3 parts MIx Diluent 0.098 P6 MIx Diluent 0.000 2 Biotinylated Albumin (1x): Dilute Biotinylated Albumin with 4 ml MIx Diluent to produce a working solution. Allow to sit for 10 minutes with gentle agitation prior to use. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate 1:20 with reagent grade water. SP Conjugate (100x): Spin down the SP Conjugate briefly and dilute the desired amount of the conjugate 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C.

    样品收集
    Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 2000 x g for 10 minutes and assay. Dilute samples 1:8000 into MIx Diluent. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. (EDTA or Heparin can also be used as anticoagulant.) Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 2000 x g for 10 minutes. Remove serum and assay. Dilute samples 1:8000 into MIx Diluent. Store serum at -20°C or below. Avoid repeated freeze-thaw cycles
    实验流程

    Prepare all reagents, working standards and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20 - 30 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccant inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. Add 25 µL of standard or sample per well, and immediately add 25 µL of Biotinylated Albumin to each well (on top of the Standard or sample) and mix gently. Cover wells with a sealing tape and incubate for one hour. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer. Invert the plate and decant the contents, and hit it 4-5 times on absorbent paper towel to complete remove liquid at each step. Add 50 µL of Streptavidin-Peroxidase Conjugate to each well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash five times with 200 µL of Wash Buffer. Add 50 µL of Chromogen Substrate per well and incubate for about 8 minutes or till the optimal blue color density develops. Gently tap plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 µL 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. Please note that after the reaction is stopped for about 10 minutes, some black particles may be generated at high concentration point, which will reduce the readings.

    结果分析

    Calculate the mean value of the 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 on the y-axis. The best-fit line can be determined by regression analysis using log-log or four-parameter logistic curve-fit. 3 Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor. Standard Curve The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

    实验精密度
    Intra-assay and inter-assay coefficients of variation were 4.6% and 7.1% respectively.
    限制
    仅限研究用
  • 注意事项
    The kit should not be used beyond the expiration date.
    储存条件
    4 °C/-20 °C
    储存方法
    Store kit at 2-8°C or -20°C upon arrival up to the expiration date. Opened MIx Diluent may be stored for up to 1 month at 2-8°C. Store reconstituted reagents at -20°C or below. Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along zip-seal. May be stored for up to 1 month in a vacuum desiccator.
  • Zhang, Yuen, Advani, Thai, Advani, Kepecs, Kabir, Connelly, Gilbert: "Early-Outgrowth Bone Marrow Cells Attenuate Renal Injury and Dysfunction via an Antioxidant Effect in a Mouse Model of Type 2 Diabetes." in: Diabetes, Vol. 61, Issue 8, pp. 2114-25, (2012) (PubMed).

    Kusinski, Stanley, Dilworth, Hirt, Andersson, Renshall, Baker, Baker, Sibley, Wareing, Glazier: "eNOS knockout mouse as a model of fetal growth restriction with an impaired uterine artery function and placental transport phenotype." in: American journal of physiology. Regulatory, integrative and comparative physiology, Vol. 303, Issue 1, pp. R86-93, (2012) (PubMed).

    Shevalye, Lupachyk, Watcho, Stavniichuk, Khazim, Abboud, Obrosova: "Prediabetic nephropathy as an early consequence of the high-calorie/high-fat diet: relation to oxidative stress." in: Endocrinology, Vol. 153, Issue 3, pp. 1152-61, (2012) (PubMed).

  • 抗原 See all Albumin (ALB) ELISA试剂盒
    Albumin (ALB)
    别名
    Albumin (ALB 产品)
    别名
    PRO0883 ELISA Kit, PRO0903 ELISA Kit, PRO1341 ELISA Kit, ALB ELISA Kit, CSA ELISA Kit, Alb-1 ELISA Kit, Alb1 ELISA Kit, Albza ELISA Kit, LOC100136344 ELISA Kit, alb-a ELISA Kit, alb-b ELISA Kit, albumin ELISA Kit, serum albumin 1 ELISA Kit, albumin S homeolog ELISA Kit, ALB ELISA Kit, Alb ELISA Kit, LOC100136344 ELISA Kit, alb.S ELISA Kit
    背景
    Albumin is a serum hepatic protein, the most abundant protein in serum and contributes to the maintenance of oncotic pressure as well as to transport of hydrophobic molecules. Serum albumin level has been linked in clinical practice to several diseases. Low albumin levels can suggest liver , kidneys disease , inflammation , shock , and malnutrition. On the other hand, high albumin levels usually reflect dehydration.
    途径
    Lipid Metabolism
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