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Laboratory Research Human ELISA Kit YKL-40 CHI3L1 Enzyme - Linked Immunosorbent Assay Kit Customized

Categories Human ELISA Kit
Brand Name: BT Lab
Model Number: Cat.No E2063Hu
Certification: CE, ISO9001:2005, MSDS
Place of Origin: Shanghai, China
MOQ: Negotiation
Price: Negotiation
Supply Ability: Western Union, T/T
Delivery Time: 1-3 business days, bulk order within one week
Packaging Details: Wrapped with ice pack and styrofoam package
Sensitivity: 0.61ng/ml
Standard Curve Range: 1ng/ml - 400ng/ml
Size: 96 wells
Shipping: DHL/FedEX
Bulk Order: Yes
Uniprot No.: P36222
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    Laboratory Research Human ELISA Kit YKL-40 CHI3L1 Enzyme - Linked Immunosorbent Assay Kit Customized

    Laboratory Research Human YKL-40 CHI3L1 Enzyme-linked Immunosorbent Assay Kit Customized


    Cat.No E2063Hu

    Storage: Store the reagents at 2-8°C. For over 6-month storage refer to the expiration date keep it at -20°C. Avoid repeated thaw cycles. If individual reagents are opened it is recommended that the kit be used within 1 month.

    *This product is for research use only, not for use in diagnosis procedures. It’s highly recommend to read this instruction entirely before use.


    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-YKL-40/CHI3L1 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.

    Intended Use

    This sandwich kit is for the accurate quantitative detection of human Chitinase-3-like Protein 1 (also known as YKL-40/CHI3L1) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.


    Specimen Collection

    Serum Allow serum to clot for 10-20 minutes at room temperature. Centrifuge at 2000-3000 RPM for 20 minutes.


    Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C within 30 minutes of collection.


    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 and other body fluids 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.


    Note

    • Sample concentrations should be predicted before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their particular experiments.
    • Samples to be used within 5 days should be stored at 2-8°C. Samples should be aliquoted or must be stored at -20°C within 1 month or -80°C within 6 months. Avoid repeated freeze thaw cycles.
    • Samples should be brought to room temperature before starting the assay.
    • Centrifuge to collect sample before use.
    • Samples containing NaN3 can’t be tested as it inhibits the activity of Horse Radish Peroxidase (HRP).
    • Collect the supernatants carefully. When sediments occurred during storage, centrifugation should be performed again.
    • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.

    *Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample matrix interference may falsely depress the specificity and accuracy of the assay.


    Reagent Preparation

    All reagents should be brought to room temperature before use.

    Standard Reconstitute the 120μl of the standard (480ng/ml) with 120μl of standard diluent to generate a 240ng/ml 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 (240ng/ml) 1:2 with standard diluent to produce 120ng/ml, 60ng/ml, 30ng/ml and 15ng/ml solutions. Standard diluent serves as the zero standard(0 ng/ml). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:


    240ng/mlStandard No.5120μl Original Standard + 120μl Standard Diluent
    120ng/mlStandard No.4120μl Standard No.5 + 120μl Standard Diluent
    60ng/mlStandard No.3120μl Standard No.4 + 120μl Standard Diluent
    30ng/mlStandard No.2120μl Standard No.3 + 120μl Standard Diluent
    15ng/mlStandard No.1120μl Standard No.2 + 120μl Standard Diluent

    Standard ConcentrationStandard No.5Standard No.4Standard No.3Standard No.2Standard No.1
    480ng/ml240ng/ml120ng/ml60ng/ml30ng/ml15ng/ml

    Wash Buffer Dilute 20ml of Wash Buffer Concentrate 25x 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.


    Calculation of Result

    Construct a standard curve by plotting the average OD for each standard on the vertical (Y) axis against the concentration on the horizontal (X) axis and draw a best fit curve through the points on the graph. These calculations can be best performed with computer-based curve-fitting software and the best fit line can be determined by regression analysis.

    Quality Laboratory Research Human ELISA Kit YKL-40 CHI3L1 Enzyme - Linked Immunosorbent Assay Kit Customized for sale
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