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96 Wells Size Specificity ELISA Test Kit , Horse Interleukin 1 Beta ELISA Kit

Categories ELISA Test Kit
Brand Name: BT Lab
Model Number: Cat.No E0079Ho
Certification: CE, ISO9001:2005, MSDS
Place of Origin: Shanghai, China
MOQ: 1 kit
Price: Negotiation
Payment Terms: Western Union, T/T
Supply Ability: In Stock
Delivery Time: 1-3 business days, bulk order within one week
Packaging Details: Wrapped with ice pack and styrofoam package
Target Protein: Interleukin 1 Beta
Standard Curve Range: 0.2ng/L - 70ng/L
Assay Time: 2 hours
Discount: Available
Customized: Acceptable
Assay Principle: Sandwich
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    96 Wells Size Specificity ELISA Test Kit , Horse Interleukin 1 Beta ELISA Kit

    Horse Interleukin 1 Beta ELISA Kit with High Spesificity and Sensitivity
    Cat.No E0079Ho
    Standard Curve Range: 0.2ng/L - 70ng/L
    Sensitivity: 0.091ng/L
    Size: 96 wells

    Reagent Provided

    ComponentsQuantity
    Standard Solution (80ng/L)0.5ml x1
    Pre-coated ELISA Plate12 * 8 well strips x1
    Standard Diluent3ml x1
    Streptavidin-HRP6ml x1
    Stop Solution6ml x1
    Substrate Solution A6ml x1
    Substrate Solution B6ml x1
    Wash Buffer Concentrate (30x)20ml x1
    Biotinylated Horse IL-1 Beta Antibody1ml x1
    User Instruction1
    Plate Sealer2 pics
    Zipper bag1 pic



    Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for approximately 20 minutes. When collecting pleuroperitoneal fluid and cerebrospinal fluid, please follow the procedures above-mentioned.

    Cell Culture Supernatant Collect by sterile tubes when examining secrete components. Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect the supernatants carefully. When examining the components within the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the cell concentration of approximately 1 million/ml. Damage cells through repeated freeze-thaw cycles to let out the inside components. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

    Tissue Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

    Reagent Preparation
    All reagents should be brought to room temperature before use.
    Standard Reconstitute the 120μl of the standard (80ng/L) with 120μl of standard diluent to generate a 40ng/L standard stock solution. Allow the standard to sit for 15 mins with gentle agitation prior to making dilutions. Prepare duplicate standard points by serially diluting the standard stock solution (40ng/L) 1:2 with standard diluent to produce 20ng/L, 10ng/L, 5ng/L and 2.5ng/L solutions. Standard diluent serves as the zero standard(0 ng/L). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:

    40ng/LStandard No.5120μl Original Standard + 120μl Standard Diluent
    20ng/LStandard No.4120μl Standard No.5 + 120μl Standard Diluent
    10ng/LStandard No.3120μl Standard No.4 + 120μl Standard Diluent
    5ng/LStandard No.2120μl Standard No.3 + 120μl Standard Diluent
    2.5ng/LStandard No.1120μl Standard No.2 + 120μl Standard Diluent

    Standard ConcentrationStandard No.5Standard No.4Standard No.3Standard No.2Standard No.1
    80ng/L40ng/L20ng/L10ng/L5ng/L2.5ng/L


    Wash Buffer Dilute 20ml of Wash Buffer Concentrate 30x into deionized or distilled water to yield 500 ml of 1x Wash Buffer. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved.

    Assay Procedure
    1. Prepare all reagents, standard solutions and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature.
    2. Determine the number of strips required for the assay. Insert the strips in the frames for use. The unused strips should be stored at 2-8°C.
    3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution contains biotinylated antibody.
    4. Add 40μl sample to sample wells and then add 10μl anti-IL-1 Beta antibody to sample wells, then add 50μl streptavidin-HRP to sample wells and standard wells (Not blank control well). Mix well. Cover the plate with a sealer. Incubate 60 minutes at 37°C.
    5. Remove the sealer and wash the plate 5 times with wash buffer. Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1 minute for each wash. For automated washing, aspirate all wells and wash 5 times with wash buffer, overfilling wells with wash buffer. Blot the plate onto paper towels or other absorbent material.
    6. Add 50μl substrate solution A to each well and then add 50μl substrate solution B to each well. Incubate plate covered with a new sealer for 10 minutes at 37°C in the dark.
    7. Add 50μl Stop Solution to each well, the blue color will change into yellow immediately.
    8. Determine the optical density (OD value) of each well immediately using a microplate reader set to 450 nm within 10 minuets after adding the stop solution.

































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