CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) Rattus norvegicus (Rat) Sandwich CLIA

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

  • CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) Packages (Simulation)
  • CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) Packages (Simulation)
  • CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) Results demonstration
  • SCG945Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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(%)
serum(n=5) 85-99 95
EDTA plasma(n=5) 87-96 92
heparin plasma(n=5) 96-105 101

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
serum(n=5) 82-101% 78-101% 81-104% 92-101%
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.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

CLIA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1)

Test principle

The microplate provided in this kit has been pre-coated with an antibody specific to Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1). Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1). 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.;

Reference

  • J Neurol SciChanges of ubiquitin C-terminal hydrolase-L1 levels in serum and urine of patients with white matter lesionsPubMed: 26232084
  • Journal of the Neurological SciencesIncreased plasma UCH-L1 after aneurysmal subarachnoid hemorrhage is associated with unfavorable neurological outcomePubmed:26810533