OxHDL ELISA Kit (Oxidized High-Density Lipoprotein)

INTENDED USE

Human OxHDL ELISA kit can measure concentrations of oxidized high-density lipoprotein (OxHDL) present is plasma, serum, biological fluids or tissue homogenate samples.

INTRODUCTION

Oxidized high-density lipoprotein (OxHDL) refers to HDL particles that have undergone oxidative modification rendering loss or alteration of normal biological activities that maintain cardiovascular health. Under healthy homeostasis, HDL transports cholesterol to liver for excretion while exerting vasoprotective antioxidant and anti-inflammatory effects that together help curb atherosclerosis. However, in pathological contexts like diabetes and cardiovascular disease, oxidative stress drives chemical changes to HDL apolipoproteins and lipids that compromise or invert functionalities enabling vascular dysfunction.

In healthy vessels, HDL particles circulate absorbing oxidation products that would otherwise damage LDL and endothelial cells. However in disease states, myeloperoxidase, 15-lipoxygenase and other reactive oxygen species override this antioxidant capacity by oxidizing both protein and lipid moieties. Consequently, proteolytic cleavage or conformational changes to apoA-I apolipoprotein in OxHDL impairs cholesterol efflux capacity from vessel walls. Furthermore, lipid peroxidation can directly deplete OxHDL’s cargo of hydrophobic antioxidants transferred to maintain redox balance. Thereby, oxidative erosion of structural and antioxidant properties promotes atherogenic effects.

Beyond passive dysfunction, OxHDL elicits endothelial apoptosis, smooth muscle migration, prothrombotic changes and inflammation activity that actively advance atherosclerotic plaque growth. This results from modifications disrupting ligand-receptor signaling affecting processes like nitric oxide production that manage vascular health and inhibition of inflammatory transcription factors. Clinically, higher circulating OxHDL levels independently predict risks for cardiovascular events, suggesting utility as prognostic biomarker. Targeting the chemical variability introduced by HDL oxidation continues to motivate drug development efforts to block pathological signaling while restoring vasoprotective activities.

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- OxHDL.
• 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 OxHDL ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 1.562, 3.125, 6.25, 12.5, 25, 50, 100 ng/ml.
Reactivity: Human
Sensitivity: 0.0938 ng/ml
Range: 0.156 – 100 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for OxHDL, no cross reactivity or interference between OxHDL and analogues was detected.
– Recovery: Serum (86 – 101%), EDTA Plasma (95 – 102%), Heparin Plasma (90 – 99%).
– Linearity: Serum (86 – 105%), EDTA Plasma (86 – 94%), Heparin Plasma (80 – 84%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Oxidized high-density lipoprotein is associated with increased plasma glucose in non-diabetic dyslipidemic subjects. Clin Chim Acta. (2012) 414: 125-129. Kotani K, et al.
2. Oxidized-high-density lipoprotein induces neuron death. Exp Neurol. (2000) 161 (2): 621-30. Keller J.N., et al.
3. OxHDL controls LOX-1 expression and plasma membrane localization through a mechanism dependent on NOX/ROS/NF-κB pathway on endothelial cells. Lab Invest. (2019) 99 (3): 421-437. Pérez L., et al.
4. Structure and Dynamics of Oxidized Lipoproteins In Vivo: Roles of High-Density Lipoprotein. Biomedicines. (2021) 9 (6): 655. Itabe H., et al.

ADDITIONAL INFORMATION

• Full Name: OxHDL ELISA Kit (Oxidized High-Density Lipoprotein)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 0.0938 ng/ml

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