Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) BioAssay(TM) ECL Kit (Rat)

Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) BioAssay(TM) ECL Kit (Rat) is a sandwich Electrochemical luminescence-based immunoassay (ECL) suitable for use in Tissue homogenates and other biological fluids.\n\nTest Principle:\nThe microtiter plate provided in this kit has been pre-coated with an antigen. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated secondary antibody. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) level in the sample or standard.\n\nDetection Range:\n78.125-5000pg/ml\n\nSensitivity:\nThe minimum detectable dose of this kit is typically less than 25pg/ml.\n\nSpecificity:\nThis sandwich assay has high sensitivity and excellent specificity for detection of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1). No significant cross-reactivity or interference between Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) and analogues was observed. \n\nStability:\nThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.\n\nPrecision:\nIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level were tested 20 times on one plate, respectively. \nInter-assay Precision (Precision between assays): 3 samples with low, middle and high level were tested on 3 different plates, 8 replicates in each plate. \n CV(%) = SD/meanX100\n Intra-Assay: CV<10%\n Inter-Assay: CV<12%\n\nKit Components:\nMicrotiter Plate, 1x96 wells, Pre-coated, ready to use,пїЅwell strip plate\nPlate sealer for 96 wells\nStandard, 2x1 vial\nStandard Diluent, 1x20ml\nDetection Reagent A, 1x120ul\nDetection Reagent B, 1x120ul\nAssay Diluent A, 1x12ml\nAssay Diluent B, 1x12ml\nSubstrate A, 1x10ml\nSubstrate B, 1x2ml\nWash Buffer, 30X, 1x20ml\n\nMaterials Required but not Supplied:\n1. Luminometer capable of reading 96-well microplates with the following parameters:пїЅlag time 30.0secs; read time 1.0 sec/well .\n2. Precision single or multi-channel pipettes and pipette tips with disposable tips.пїЅ\n3. Eppendorf Tubes for diluting samples.\n4. Deionized or distilled water.\n5. Absorbent paper for blotting the microtiter plate.\n6. Container for Wash Solution\n\nProtocol Summary:\n1. Prepare all reagents, samples and standards.\n2. Add 100ul standard or sample to each well. Incubate 2 hours at 37 degrees C.\n3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37 degrees C.\n4. Aspirate and wash 3 times.\n5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37 degrees C.\n6. Aspirate and wash 5 times.\n7. Add 100ul Substrate Solution. Incubate 10 minutes at 37 degrees C.\n8. Read RLU value immediately.\n\nSample Collection:\nSerum:\nUse a serum separator tube and allow samples to clot for two hours at RT or overnight at 4 degrees C before centrifugation for 20 minutes at approximately 1000пїЅg. Assay freshlyпїЅprepared serumпїЅimmediately or store samples in aliquot at -20 degrees C or -80 degrees C for later use. Avoid repeated freeze/thaw cycles.\n\nPlasma:\nCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes atпїЅ1000пїЅg at 2-8 degrees C within 30 minutes of collection.\nRemove plasma and assay immediatelyпїЅor store samplesпїЅin aliquots at -20 degrees C or -80 degrees C for later use. Avoid repeated freeze/thaw cycles.\n\nOther Biological Fluids:\nCentrifuge samples for 20 minutes at 1000xg. Remove particulates and assayпїЅimmediately or store samples in aliquot at -20 degrees C or -80 degrees C for later use.\nAvoid repeatedпїЅfreeze/thaw cycles.\n\nNote:\nSamples to be used within 5 days may be stored at 4 degrees C, otherwise samples must be stored at -20 degrees C ( 1пїЅmonth) or -80 degrees C ( 2 months) to avoid loss of bioactivity and contamination.\nSample hemolysis will influence the result, so hemolytic specimens cannot be detected.\nWhen performing the assay, bring samples to room temperature.\n\nStorage and Stability:\n1. For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degrees C upon receipt while the others should be at 4 degrees C.\n2. For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.\n3. Expiration date is 6 months from shipment date.\n4. It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.

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SPECIFICATIONS

Catalog Number

158185

Size

96Tests

Additional Info

This sandwich assay has high sensitivity and excellent specificity for detection of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1). No significant cross-reactivity or interference between Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) and analogues was observed.