Galectin-9 ELISA Kit (GAL9)

Full Name: Galectin-9 ELISA Kit (GAL9)
Reactivity: Human
Sample Type: Tissue Homogenates, Plasma, Serum, Biological Fluids
Sensitivity: 0.094 ng/ml

INTRODUCTION

Galectin-9 (GAL9) can be phosphorylated on serine residues by casein kinase 1 (CK1). Phosphorylation can also occur due to protein kinase A or phosphatidylin ositol-3-kinase. An increase in the intracellular calcium concentration by the binding of phosphorylated GAL-9 to the cell’s ER membrane results in the release of calcium from the endoplasmic reticulum and the proteolysis of membrane proteins. It can also be cleaved by thrombin and casein kinase 1. Cleavage results in an activation of the protein’s signalling mechanism, which can trigger a proteolytic cascade that leads to cell death. It is involved in cell adhesion, and it induces the expression of integrins α4 and β1, which are involved in migration.

Galectin-9 facilitates adhesion via the interaction of the lectin domains with different molecules on the cell surface. It is found to exhibit a high affinity for collagen and fibronectin (GFN). The binding to GFN results in changes that modulate cell migration and proliferation, allowing cells to adhere more strongly. It interacts with members of the integrin family, which are proteins involved in maintaining cell adhesion. It binds to α4β1 integrins and their combination increases adhesive properties in focal adhesions, the sites of cell-to-cell adhesion. This binding creates an adhesive zone that is 25% more adhesive than control cells.

GAL9 has important cytoplasmic, intracellular functions and controls AMPK in response to lysosomal damage that can occur upon exposure to endogenous and exogenous membrane damaging agents such as crystalline silica, cholesterol crystals, microbial toxins, proteopathic aggregates such as tau fibrils and amyloids. It is one of the most studied ligands for HAVCR2 (TIM-3) and is expressed on various tumor cells. It can also interact with other proteins such as CLEC7A, CD137, CD401. For example, an interaction with CD40 on T-cells induced their proliferation inhibition and cell death.

INTENDED USE

Human galectin-9 ELISA kit can detect amounts GAL9 (galectin-9, LGALS9, Eecalectin, HUAT) present is human serum, plasma, biological fluids and tissue homogenate samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with anti-GAL9 antibody.
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

For this galectin-9 ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 10 ng/ml.
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for GAL9, no cross reactivity or interference between GAL9 and analogues was detected.
– Recovery: Serum (86 – 99%), EDTA Plasma (88 – 101%), Heparin Plasma (92 – 99%).
– Linearity: Serum (95 – 101%), EDTA Plasma (82 – 98%), Heparin Plasma (84 – 98%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

Galectin-9: A Suppressor of Atherosclerosis? Front Immunol. (2020) 11: 604265. Yu J., et al.
The TIM3/Gal9 signaling pathway: An emerging target for cancer immunotherapy. Cancer Lett. 2021 Jul 10;510:67-78. Kandel S., et al.
Endometrial regenerative cells with galectin-9 high-expression attenuate experimental autoimmune hepatitis. Stem Cell Res Ther. 2021 Oct 15;12(1):541. Wang H., et al.

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