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Multiplex Assay Kit for Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay)

CPSD; CLN10; Lysosomal Aspartyl Protease; Ceroid-Lipofuscinosis,Neuronal 10

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-101 97
EDTA plasma(n=5) 87-101 95
heparin plasma(n=5) 93-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathepsin D (CTSD) ,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 Cathepsin D (CTSD) ,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 Cathepsin D (CTSD) ,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) 79-93% 86-94% 79-95% 96-104%
EDTA plasma(n=5) 86-101% 78-102% 99-105% 80-93%
heparin plasma(n=5) 89-102% 91-99% 82-95% 88-99%

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#:CTSD) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Molecular Immunology Cathepsin D is released after severe tissue trauma in vivo and is capable of generating C5a in vitro ScienceDirect: S0161589011008297
21 Plasma Cathepsin D Levels: A Novel Tool to Predict Pediatric Hepatic Inflammation PubMed: 25732418
Scientific Reportes Plasma cathepsin D correlates with histological classifications of fatty liver disease in adults and responds to intervention pubmed:27922112
Biomedicine & pharmacotherapy Berberine ameliorates intrahippocampal kainate-induced status epilepticus and consequent epileptogenic process in the rat: Underlying mechanisms pubmed:28061403
JGH OPEN Limited applicability of cathepsin D for the diagnosis and monitoring of non‐alcoholic steatohepatitis
Diabetologia Plasma cathepsin D activity is negatively associated with hepatic insulin sensitivity in overweight and obese humans Pubmed: 31690989
Antioxidants Iron and Advanced Glycation End Products: Emerging Role of Iron in Androgen Deficiency in Obesity Pubmed: 32235809
Peptides LVV-hemorphin-7 (LVV-H7) plays a role in antinociception in a rat model of alcohol-induced pain disorders 33253777
Cells Identification of Cathepsin D as a Plasma Biomarker for Alzheimer's Disease 33445607
J Chem Neuroanat Analysis of silymarin-modulating effects against acrylamide-induced cerebellar damage in male rats: Biochemical and pathological markers 33965515
Front Cell Infect Microbiol PGRS Domain of Rv0297 of Mycobacterium tuberculosis Is Involved in Modulation of Macrophage Functions to Favor Bacterial Persistence 33042856
Nutr Metab Cardiovasc Dis Serum CathepsinD in pregnancy: relation with metabolic and inflammatory markers and effects of fish oils and probiotics Pubmed:35304048
Biosensors A Comparison of Various Chips Used for the Manufacture of Biosensors Applied in Non-Fluidic Array SPRi, Based on the Example of Determination of Cathepsin D Pubmed:35049649
Catalog No. Related products for research use of Bos taurus; Bovine (Cattle) Organism species Applications (RESEARCH USE ONLY!)
RPB280Bo01 Recombinant Cathepsin D (CTSD) Positive Control; Immunogen; SDS-PAGE; WB.
PAB280Bo01 Polyclonal Antibody to Cathepsin D (CTSD) WB; IHC; ICC; IP.
MAB280Bo21 Monoclonal Antibody to Cathepsin D (CTSD) WB; IHC; ICC; IP.
SEB280Bo ELISA Kit for Cathepsin D (CTSD) Enzyme-linked immunosorbent assay for Antigen Detection.
CEB280Bo ELISA Kit for Cathepsin D (CTSD) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB280Bo Multiplex Assay Kit for Cathepsin D (CTSD) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.