Sandwich ELISA is a widely used immunoassay technique designed for detecting and quantifying specific target proteins in various sample types. Unlike direct or indirect ELISAs, sandwich ELISA offers superior specificity by using two antibodies that bind to different epitopes on the target protein. Our catalog includes quantitative, semi-quantitative, and qualitative assays, with innovative SimpleStep ELISA® kits that reduce assay time to 90 minutes while maintaining our signature high specificity and performance. These kits utilize highly specific matched antibody pairs, ensuring sensitive and consistent detection of target analytes, making them ideal for high-throughput screening (HTS) and various research applications.
FEATURES
- High specificity: Utilizes two antibodies targeting different epitopes on the same antigen.
- Suitable for complex samples: Effectively detects targets in heterogeneous mixtures.
- High flexibility and sensitivity: Supports both direct and indirect detection methods.
- Short assay times: Typically, 1.5-2.5 hours with broad detection ranges.
- Fast and easy protocol: Our SimpleStep ELISA® features a single-wash procedure, reducing assay time to 90 minutes or less.
- Versatile and validated: Uses recombinant monoclonal antibodies, validated in various biological samples, and available in multiple formats.
APPLICATIONS
- Quantitative analysis: Accurately measures protein concentrations in biological samples.
- Semi-quantitative detection: Allows for relative protein quantification.
- Qualitative assessment: Identifies the presence of target molecules.
- HTS: Enables large-scale drug discovery and biomarker validation using 96- and 384-well plate formats.
FAQs
What is a Sandwich ELISA, and how does it work?
A Sandwich ELISA is a highly specific immunoassay that detects and quantifies target proteins using two antibodies that bind to distinct epitopes. A capture antibody immobilized on a plate first binds the target protein from a sample, followed by a detection antibody that forms a sandwich complex. An enzyme-linked reaction then generates a measurable signal with intensity proportional to the protein concentration.
How do I choose the right capture and detection antibodies for a Sandwich ELISA?
Ensure the antibodies bind to different, non-overlapping epitopes on the target protein to avoid competitive binding. Use a high-affinity recombinant monoclonal antibody as the capture antibody for strong and specific target binding and consistent results. The detection antibody should also be highly specific and can be either directly enzyme-conjugated or detected via a secondary antibody. Abcam offers kits with matched antibody pairs with confirmed distinct epitopes, pre-validated for ELISA, that offer the best reliability and reproducibility.
What factors affect the sensitivity and specificity of a Sandwich ELISA?
The sensitivity and specificity depend on factors such as antibody quality, epitope selection, and sample integrity. High-affinity, well-matched antibody pairs improve target binding, while proper blocking, washing, and detection methods minimize background noise. Optimized incubation conditions and plate coating efficiency enhance assay reproducibility and signal strength.