Traditional ELISA Kits

Traditional ELISA (enzyme-linked immunosorbent assay) kits are a common laboratory technique used to detect and quantify the amount of analyte in a given sample.

Reddot Biotech's Traditional ELISA kits are our foundational ELISA kit that has a conventional workflow at a reasonable price. These kits come with pre-coated plates and all the reagents you need to perform the experiment. With a simple protocol and easy to follow instructions, these kits are the perfect starting point, while maintaining the same high level of quality and sensitivity.

Browse Traditional ELISA Kits

Key Features & Benefits

• Wide Variety of Targets - over 6500 different kits.
• Complete - includes pre-coated plates and everything you need except your samples.
• High Sensitivity & Specificity - Detects low-abundance analytes with accuracy.
• Versatile Applications - Compatible with many different sample types.
• Many different reactivities available -including hard to find species.
• Straightforward procedure - easy-to-follow steps.
• Affordable - prices to fit all budgets including large projects.
• Reliable & Reproducible Results - Optimized reagents for consistent performance.

Types of ELISA Kits

Reddot Biotech offers two formats of Traditional ELISA kits:

Sandwich ELISA

• Uses two antibodies specific to the target analyte
• High sensitivity and specificity
• Commonly used for cytokine and protein detection

How a Sandwich ELISA works:

When using a sandwich ELISA kit, the microtiter plate included in the kit is pre-coated with an antibody specific to the target of interest. After the samples are prepared, samples and standards are added to the plate wells with a biotin-conjugated antibody preparation specific to the target. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Next, a TMB substrate is added to bind to the HRP. After the TMB substrate solution addition, only those wells that contain the target, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 ± 10 nm. The concentration of the target in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Competitive Inhibition ELISA

• Measures small molecules, hormones, and metabolites
• High sensitivity, even for low-abundance targets
• Ideal for detecting molecules with a single binding site

How a Competitive Inhibition ELISA works:

In the case of competitive inhibition ELISA kits, a competitive inhibition reaction is launched between biotin labeled target and unlabeled target (Standards or samples) with the pre-coated antibody (or occasionally antigen)  specific to that target. After incubation, the unbound conjugate is washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is inversely proportional to the concentration of target in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of the target in the sample.

How to Choose the Right ELISA Kit

Make sure you choose the right ELISA kit for you by considering the following:

• Target Analyte - Identify whether you are detecting a protein, antibody, or small molecule.
• Sample Type Compatibility - Ensure the kit is optimized for your sample (e.g., serum, plasma, tissue lysates).
• Sensitivity & Dynamic Range - Consider the lowest detection limits required for your research.

If you need help choosing the right kit for you, or require a custom manufactured kit, please contact us here for more information.

• Pre-coated 96-well strip plate
• 2 Plate sealers for 96 wells 
• 2 Standards
• Diluent Buffer
• Wash Buffer (30× concentrate)
• Detection Reagent A  
• Detection Reagent B 
• TMB Substrate 
• Stop Solution 
• Instruction manual

Traditional ELISA Kit Procedure:

1. Prepare all reagents, samples and standards.
2. Add 100 μL standard or sample to each well. Incubate 2 hours at 37°C.
3. Aspirate and add 100 μL prepared Detection Reagent A. Incubate 1 hour at 37°C.
4. Aspirate and wash 3 times.
5. Add 100 μL prepared Detection Reagent B. Incubate 1 hour at 37°C.
6. Aspirate and wash 5 times.
7. Add 90 μL Substrate Solution. Incubate 15-25 minutes at 37°C.
8. Add 50 μL Stop Solution. Read at 450 nm immediately.

Frequently Asked Questions (FAQs)

Q: How are Traditional and Ready-to-Use ELISA kits different?
A: Ready-to-Use ELISA kits have some slight optimizations to make them more user-friendly, but our Traditional kits have a lower price point. Both options have the same high level of quality and sensitivity.

Q: How should I store my Traditional ELISA kits?
A: Always store kits as directed in the manual or on labels. For Traditional ELISA kits from Reddot Biotech, store the TMB Substrate, Wash Solution Concentrate, and Stop Solution at 4^oC, and the rest of the components at -20^oC for up to 12 months.

Q: What dilution should I use for my samples with Traditional ELISA kits?
A: The dilution for your samples will depend on the concentration of the analyte in your sample. For best results, we recommend performing a preliminary concentration gradient experiment to determine the optimal dilution ratio for your samples before you begin. For reference values or more information, contact us here.

Explore Our ELISA Kit Selection

Browse our comprehensive range of Traditional ELISA Kits to find the best option for your research needs. Need assistance? Contact our support team for expert guidance, or explore our other ELISA kit ranges here:

Ready-To-Use ELISA Kits

EASYStep Pro ELISA Kits

Food Safety ELISA Kits