CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1)

PARK5; PGP9.5; Uch-L1; Neuron cytoplasmic protein 9.5; Ubiquitin thioesterase L1; Ubiquitin carboxyl-terminal hydrolase isozyme L1
Typical Standard Curve Typical Standard Curve

Sample type Serum, plasma, tissue homogenates, cell lysates and other biological fluids. Detection range 20.576-15000pg/mL The standard curve concentrations used for the ELISA's were 15000pg/mL, 5000pg/mL, 1666.67pg/mL, 555.56pg/mL, 185.185pg/mL, 61.728pg/mL, 20.576pg/mL
Sensitivity The minimum detectable dose of this kit is typically less than 7.3pg/mL. Specificity This 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.

Recovery Matrices listed below were spiked with certain level of recombinant Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) and the recovery rates were calculated by comparing the measured value to the expected amount of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) in samples.
Matrix Recovery range (%) Average(%)
EDTA plasma(n=5)87-9692
heparin plasma(n=5)96-105101

Precision Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample 1:2 1:4 1:8 1:16
EDTA plasma(n=5)86-101%85-95%89-102%86-101%
heparin plasma(n=5)83-101%85-101%85-92%87-97%

Stability The 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.
Test principle The 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. Assay procedure summary 1. Prepare all reagents, samples and standards;
Reagent Preparation
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 100µL Substrate Solution. Incubate 10 minutes at 37oC;
8. Read RLU value immediately.

Reagents and materials provided
  • Pre-coated, ready to use 96-well strip plate1
  • Plate sealer for 96 wells4
  • Standard2
  • Standard Diluent1×20mL
  • Detection Reagent A1×120µL
  • Assay Diluent A1×12mL
  • Detection Reagent B1×120µL
  • Assay Diluent B1×12mL
  • Substrate A1×10mL
  • Substrate B1×2mL
  • Wash Buffer (30 × concentrate)1×20mL
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