Human Glycated Low-Density Lipoprotein (Glycated-LDL) ELISA Kit

The Human Glycated LDL ELISA Kit is a high-sensitivity, sandwich-based ELISA assay designed for the quantitative detection of glycated low-density lipoprotein (Glycated-LDL) in human serum, plasma, cell culture supernatants, tissue homogenates, and body fluids. This kit is an essential research tool for scientists studying glycation-associated oxidative stress, cardiovascular complications, and metabolic disorders, particularly in diabetes-related lipid dysfunction.

Key Features

• Assay Type: Sandwich ELISA
• Detection Range: 0.06 – 4 ng/mL
• Sensitivity: 0.01 ng/mL
• Sample Volume: 10 μL
• Assay Time: ~2.5 hours
• Storage: 2–8 °C
• Shelf Life: 6 months
• Precision:
  • Intra-assay CV <10%
  • Inter-assay CV <12%

Assay Principle

This ELISA uses a sandwich-based format where a microplate is pre-coated with an anti-Glycated-LDL antibody. Upon the addition of standards or samples, a HRP-conjugated detection antibody binds the target. After washing, TMB substrate is added to develop a blue color, which turns yellow with stop solution. The color intensity (measured at 450 nm) is directly proportional to the concentration of Glycated-LDL, allowing quantification using a standard curve.

Kit Components

Sample Types & Preparation

Validated for:

• Serum / Plasma: Standard centrifugation (2,000–3,000 rpm, 20 min).
• Urine / Body Fluids / CSF: Collected and centrifuged as above.
• Cell Supernatants: Collect and clarify by centrifugation.
• Tissue Homogenates: Homogenize in PBS (pH 7.4), centrifuge, and collect supernatant.

Note: Avoid sodium azide (NaN₃) and repeated freeze-thaw cycles. Samples can be stored at –20 °C if not used immediately.

Assay Procedure (Overview)

1. Prepare standard dilutions from 5.4 ng/mL down to 0.3 ng/mL.
2. Add 40 μL sample diluent + 10 μL sample per well (except blank).
3. Incubate for 30 minutes at 37 °C.
4. Wash plate 5 times with diluted wash buffer.
5. Add 50 μL HRP-conjugated antibody; incubate 30 minutes.
6. Wash again, add 50 μL each of Chromogen A and B; incubate 15 minutes (dark).
7. Stop reaction with 50 μL stop solution.
8. Read OD at 450 nm within 15 minutes.

Performance & Precision

Standard curve plotting is performed on a log-log scale. Sample concentrations are calculated based on OD values and multiplied by the sample dilution factor.

The Human Glycated-LDL ELISA Kit provides a powerful, reproducible, and sensitive platform for researchers examining glycation-modified lipoproteins, an important biomarker in atherosclerosis, diabetes, and metabolic disorders. With a broad detection range, low sample volume requirement, and validated performance across multiple sample types, this kit offers high reliability and convenience for basic and translational research in lipid biochemistry and chronic disease models.