ELISA Kit for Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-UCHL1)

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

Specificity

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

Recovery

Matrices listed below were spiked with certain level of Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-UCHL1) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-UCHL1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-95 92
EDTA plasma(n=5) 95-105 101
heparin plasma(n=5) 78-101 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-UCHL1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-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 Anti-Ubiquitin Carboxyl Terminal Hydrolase L1 Antibody (Anti-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) 94-102% 84-95% 87-104% 80-97%
EDTA plasma(n=5) 88-95% 84-101% 88-101% 78-105%
heparin plasma(n=5) 96-105% 96-105% 87-94% 97-105%

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
TMB Substrate 1×9mL Stop Solution 1×6mL
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 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
J Neurol Sci Changes of ubiquitin C-terminal hydrolase-L1 levels in serum and urine of patients with white matter lesions PubMed: 26232084
Journal of the Neurological Sciences Increased plasma UCH-L1 after aneurysmal subarachnoid hemorrhage is associated with unfavorable neurological outcome Pubmed:26810533
Ulutas Medical Journal Serum Carnosine Dipeptidase 1 and Ubiquitin C - Terminal Hydrolase L1 as Markers of Brain Damage in Patients after Carotid Endarterectomy mnstemps:135
Scientific Reports Activation of hepatic stellate cells by the ubiquitin C-terminal hydrolase 1 protein secreted from hepatitis C virus-infected hepatocytes pubmed:28667290
Intensive Care Medicine Relationships between markers of neurologic and endothelial injury during critical illness and long-term cognitive impairment and disability Pubmed:29523900
Earth and Environmental Science Serum concentration of ubiquitin c-terminal hydrolase-L1 in detecting severity of traumatic brain injury article:10.1088
Peripheral blood biomarkers in aneurysmal subarachnoid hemorrhage ISBN:978-952-03-0750-9
Journal of Critical Care Association of neuronal repair biomarkers with delirium among survivors of critical illness Pubmed: 31896448
Brain Sciences BDNF and IL-8, But Not UCHL-1 and IL-11, Are Markers of Brain Injury in Children Caused by Mild Head Trauma Pubmed: 32987792
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