Biotin ELISA Kit - 10 x 96 Tests

Overview of Biotin ELISA Kit

The Biotin ELISA Kit is a highly sensitive and specific enzyme-linked immunosorbent assay designed to quantitatively detect biotin levels in biological samples. It operates on the principle of antigen-antibody interaction, where biotin (the target antigen) is captured by specific antibodies coated onto the wells of a microplate. This process allows for the detection of biotin with high precision.

Components:

1. Coated Wells: Microplate wells pre-coated with biotin-specific antibodies.
2. Biotin Standards: Known concentrations of biotin to generate a standard curve for quantification.
3. Sample: The biological sample suspected to contain biotin.
4. Biotin Conjugate: An enzyme-labeled biotin analog that binds to the biotin in the sample, forming a biotin-antibody-enzyme complex.
5. Substrate Solution: Typically a chromogenic or fluorogenic substrate that reacts with the enzyme, producing a measurable signal (color change or fluorescence).
6. Wash Buffer: Used to remove unbound substances during the assay to reduce background interference.

Principle:

The Biotin ELISA Kit follows a sandwich ELISA technique. During the assay, biotin in the sample binds to the capture antibody immobilized on the plate. The biotinylated enzyme-conjugate binds to the captured biotin. After washing, the bound conjugate is reacted with a substrate to produce a colorimetric or fluorescent signal. The intensity of the signal is proportional to the concentration of biotin in the sample.

Steps:

1. Coating: Biotin-specific antibodies are adsorbed onto the surface of a microplate.
2. Blocking: To prevent non-specific binding, uncoated areas of the plate are blocked with a protein solution.
3. Incubation: The sample is added to the wells, followed by the biotin enzyme-conjugate. After incubation, the plate is washed to remove unbound material.
4. Substrate Reaction: A substrate is added that reacts with the enzyme, producing a color change or fluorescence.
5. Measurement: The signal is quantified using a microplate reader at the appropriate wavelength (e.g., 450 nm for colorimetric assays or 490 nm for fluorescence-based assays).

Applications:

• Biotin quantification in serum, plasma, or other biological fluids.
• Monitoring of biotin levels in nutritional studies, metabolic research, and disease diagnosis.
• Quality control in biotin-based product manufacturing.

Sensitivity and Specificity:

The Biotin ELISA Kit is designed to provide high sensitivity with a low detection limit, making it suitable for detecting even trace amounts of biotin. The specificity of the kit ensures minimal cross-reactivity with other compounds, providing reliable and accurate results.

Advantages:

• High precision and accuracy in biotin detection.
• Quantitative results that are easy to interpret.
• Minimal sample preparation required.
• Short assay time, typically within 2-3 hours.
• Wide dynamic range, suitable for both high and low concentrations of biotin.