Human low-density lipoprotein, LDL ELISA Kit

The Human LDL ELISA Kit is a high-sensitivity, sandwich-based enzyme-linked immunosorbent assay (ELISA) designed for the quantitative measurement of low-density lipoprotein (LDL) levels in a wide range of biological samples, including serum, plasma, cell culture supernatants, tissue homogenates, and other body fluids. This assay enables precise detection of LDL in as little as 10 μL of sample, making it ideal for cardiovascular research, lipid metabolism studies, and related biomedical investigations.

Key Features

• Assay Type: Sandwich ELISA
• Detection Range: 0.3 – 10 ng/mL
• Sensitivity: 0.08 ng/mL
• Sample Volume: 10 μL
• Assay Time: ~2.5 hours
• Storage Conditions: 2–8°C
• Shelf Life: 6 months
• Precision:
  • Intra-assay CV <10%
  • Inter-assay CV <12%
• Validated for: Serum, plasma, urine, cerebrospinal fluid, pleural fluid, cell culture supernatants, tissue lysates

Principle of the Assay

This assay utilizes a sandwich ELISA format. A microplate pre-coated with an anti-human LDL monoclonal antibody captures LDL from the sample. After washing, an HRP-conjugated secondary antibody binds the captured LDL. Upon the addition of TMB substrate, color develops proportionally to the amount of LDL present. The reaction is stopped, and absorbance is measured at 450 nm, allowing quantification against a prepared standard curve.

Kit Contents

Sample Types & Preparation

• Serum/Plasma: Centrifuge at 2,000–3,000 rpm for 20 minutes after clotting or collection.
• Urine/Body Fluids: Centrifuge, then collect supernatant.
• Cell Supernatants: Centrifuge post-culture. For intracellular LDL, lyse cells via freeze-thaw cycles.
• Tissue Homogenates: Homogenize with PBS (pH 7.4) at 4°C, centrifuge, and collect supernatant.

Note: Avoid sodium azide (NaN₃), which inhibits HRP. Minimize freeze-thaw cycles.

Assay Procedure (Overview)

1. Prepare serial dilutions of the LDL standard (0.75–13.5 ng/mL).
2. Add 40 μL sample diluent and 10 μL sample to wells.
3. Incubate at 37°C for 30 minutes.
4. Wash plate 5 times with diluted wash buffer.
5. Add 50 μL HRP-conjugated antibody; incubate again.
6. Repeat wash step.
7. Add 50 μL each of Chromogen A and B; incubate for 15 minutes in the dark.
8. Add 50 μL Stop Solution.
9. Read absorbance at 450 nm within 15 minutes.

Data Analysis

• Generate a standard curve by plotting OD450 against known concentrations.
• Determine sample concentrations from the curve and multiply by the dilution factor.
• Recommended to include replicates and a standard curve in each assay.

Precision and Sensitivity

The Human LDL ELISA Kit (SL1095Hu) is a robust, sensitive, and easy-to-use assay system designed for precise quantification of LDL in various human biological matrices. Its optimized protocol, high reproducibility, and compatibility with a wide range of sample types make it a reliable tool for researchers studying lipid metabolism, cardiovascular risk markers, and cholesterol-related disorders. With clear procedural steps and validated performance metrics, this kit delivers accurate results essential for translational and preclinical research settings.