Multiplex Assay Kit for Galectin 9 (GAL9) ,etc. by FLIA (Flow Luminescence Immunoassay)

LGALS9; Ecalectin; Lectin,Galactoside-Binding Soluble 9; Tumor antigen HOM-HD-21

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Galectin 9 (GAL9) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Galectin 9 (GAL9) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Galectin 9 (GAL9) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Galectin 9 (GAL9) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 95-103 99
heparin plasma(n=5) 78-90 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Galectin 9 (GAL9) ,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 Galectin 9 (GAL9) ,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 Galectin 9 (GAL9) ,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) 85-102% 79-102% 97-105% 87-96%
EDTA plasma(n=5) 99-105% 86-103% 90-98% 94-102%
heparin plasma(n=5) 91-98% 85-92% 84-101% 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#:GAL9) 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
American Journal of Nephrology Temporal Increases in Urinary Carboxymethyllysine Correlate with Albuminuria Development in Diabetes Karger: 328581
Clinical Immunology Galectin-9 administration ameliorates CVB3 induced myocarditis by promoting the proliferation of regulatory T cells and alternatively activated Th2 cells ScienceDirect: S1521661611001057
Renal Failure Rat Renal Interstitial Fibroblasts Affect the Th1/Th2 Profile In Vitro Pubmed: 22013937
BMC Nephrol Serum galectin-9 levels are elevated in the patients with type 2 diabetes and chronic kidney disease PubMed: 23339460
Retrovirology Dysregulated Tim-3 expression on natural killer cells is associated with increased Galectin-9 levels in HIV-1 infection PubMed: PMC3750478
PLoS One. Upregulation of the Tim-3/Galectin-9 Pathway of T Cell Exhaustion in Chronic Hepatitis B Virus Infection PubMed: PMC3480425
PLoS Pathogens Galectin-9 and IL-21 Mediate Cross-regulation between Th17 and Treg Cells during Acute Hepatitis C PubMed: PMC3688567
PLOS ONE Graves’ Disease Is Associated with a Defective Expression of the Immune Regulatory Molecule Galectin-9 in Antigen-Presenting Dendritic Cells PubMed: 25880730
Advances in Experimental Medicine and Biology Molecules of Damage-Associated Patterns in Bronchoalveolar Lavage Fluid and Serum in Chronic Obstructive Pulmonary Disease Pubmed:29429028
Journal of Microbiology, Immunology and Infection Galectin-9-like from Angiostrongylus cantonensis young adult worms modulates eosinophil chemotaxis in vitro Doi: 10.1016/j.jmii.2018.09.010
Oncology Letters The effects of Tim‑3 activation on T‑cells in gastric cancer progression
Scientific Reports Circulating levels of PD-L1 and Galectin-9 are associated with patient survival in surgically treated Hepatocellular Carcinoma independent of their intra … Pubmed: 31337865
Biomolecules Blood Levels of Galectin-9, an Immuno-Regulating Molecule, Reflects the Severity for the Acute and Chronic Infectious Diseases. Biomolecules 2021, 11, 430 33804076
Medicine Regulatory T cells, damage-associated molecular patterns, and myeloid-derived suppressor cells in bronchoalveolar lavage fluid interlinked with chronic obstructive … Pubmed:35687771
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPA309Hu01 Recombinant Galectin 9 (GAL9) Positive Control; Immunogen; SDS-PAGE; WB.
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EPA309Hu61 Eukaryotic Galectin 9 (GAL9) Positive Control; Immunogen; SDS-PAGE; WB.
PAA309Hu01 Polyclonal Antibody to Galectin 9 (GAL9) WB; IHC
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