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Multiplex Assay Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity for detection of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay).
100% cross-reactivity of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay) was observed among Human, Mouse, Rat.

Recovery

Matrices listed below were spiked with certain level of recombinant Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-92 88
EDTA plasma(n=5) 82-90 85
heparin plasma(n=5) 92-99 96
sodium citrate plasma(n=5) 78-98 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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% 85-98% 80-94% 98-105%
EDTA plasma(n=5) 93-101% 89-96% 91-105% 85-99%
heparin plasma(n=5) 95-105% 86-102% 91-98% 78-102%
sodium citrate plasma(n=5) 78-105% 80-98% 86-93% 78-102%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:UCHL1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
Catalog No. Related products for research use of Homo sapiens (Human), Mus musculus (Mouse), Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
SEG945Mi ELISA Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMG945Mi Multiplex Assay Kit for Ubiquitin Carboxyl Terminal Hydrolase L1 (UCHL1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.