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|Target Protein:||Meteorin-like Protein||Assay Principle:||Sandwich|
|Standard Curve Range:||0.05ng/ml - 15ng/ml||Sensitivity:||0.023ng/ml|
elisa reagent kit,
Customized Human METRNL ELISA Assay Kit Meteorin-like Protein ELISA Test Kit For Research
Size: 96 wells
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.
This ELISA kit applies to the invitro quantitative determination of human METRNL concentrations in serum, plasma, tissue and other biological fluids.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
CV(%) = SD/mean x 100
|Standard Solution (16ng/ml)||0.5ml x1|
|Pre-coated ELISA Plate||12 * 8 well strips x1|
|Standard Diluent||3ml x1|
|Stop Solution||6ml x1|
|Substrate Solution A||6ml x1|
|Substrate Solution B||6ml x1|
|Wash Buffer Concentrate (25x)||20ml x1|
|Biotinylated human METRNL Antibody||1ml x1|
|Plate Sealer||2 pics|
|Zipper bag||1 pic|
Samples should be clear and transparent and be centrifuged to remove suspended solids.
Serum: Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1000×g. Collect the supernatant and carry out the assay immediately. Blood collection tubes should be disposable, non-pyrogenic, and non-endotoxin.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2 - 8°Cwithin 30 minutes of collection. Collect the supernatant and carry out the assay immediately. Hemolysissamples are not suitable for ELISA assay!
Cell culture supernate: Centrifuge supernate for 20 minutes to remove insoluble impurity and cell debris at 1000×g at 2 - 8°C. Collect the clear supernate and carry out the assay immediately.
Tissue homogenates:You’d better get detailed references from other literatures before assay aiming at different tissue types. For general information, hemolysis blood may affect the result, so you should mince the tissues to small pieces and rinse them in ice-cold PBS (0.01M, pH=7.4) to remove excess blood thoroughly. Tissue pieces should be weighed and then homogenized in PBS (the volume depends on the weight of the tissue) with a glass homogenizer on ice. To further break the cells, you can sonicate the suspension with an ultrasonic cell disrupter or subject it to freeze-thaw cycles. The homogenates are then centrifugated for 5minutes at 5000×g to get the supernate.
Other biological fluids: Centrifuge samples for 20 minutes at 1000×g at 2 - 8°C. Collect the supernatant and carry out the assay immediately.
*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.
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-METRNL 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.
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.
Contact Person: Lee
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