Product Description
The Protein A ELISA Kit is designed for the quantitative detection of recombinant and alkali-tolerant Protein A in biological samples. This kit uses a double-antibody sandwich ELISA method, ensuring high specificity and sensitivity, with a detection limit of 0.05 ng/mL. It is ideal for separating Protein A from IgG products and is compatible with humanized monoclonal antibody concentrations up to 2 mg/mL.
Experimental Overview
- Catalog Number: ATK00052
- Detection wavelength: 450 nm
- Sensitivity: 0.05 ng/mL
- Precision: CV < 10% for concentrations > 0.3 ng/mL
- Hook Effect: Observed at 10 μg/mL
Components & Storage
| Component | Details | Storage |
|---|---|---|
| Protein A Standard | Recombinant alkali-tolerant Protein A, 50 μg/mL, 1x0.1mL | 2-8°C |
| Biotinylated Antibody | Biotinylated anti-Protein A monoclonal antibody, 1x13mL | 2-8°C |
| Sample Diluent | Tris buffered saline, 1x30mL | 2-8°C |
| Denaturing Buffer | Citrate buffer, 1x20mL | 2-8°C |
| 10x Assay Buffer | Phosphate buffer, 1x30mL | 2-8°C |
| Streptavidin-HRP | Streptavidin-labeled HRP, 1x13mL | 2-8°C |
| TMB Substrate | 3,3’,5,5’-Tetramethylbenzidine, 1x13mL | 2-8°C |
| Stop Solution | 2M sulfuric acid, 1x13mL | 2-8°C |
| 96-well Coated Plate | Anti-Protein A antibody-coated strips, 12x8 wells | 2-8°C |
Procedure Summary
- Prepare standards, samples, and reagents according to the kit instructions.
- Add 100 μL standards, samples, and controls to the wells.
- Add 50 μL Denaturing Buffer to each well, mix, and incubate at room temperature for 10 minutes.
- Add 100 μL Biotinylated Antibody to each well and incubate at room temperature for 1 hour.
- Wash the plate 4 times with diluted Wash Buffer.
- Add 100 μL Streptavidin-HRP to each well and incubate for 10 minutes.
- Wash the plate 4 times, then add 100 μL TMB Substrate and incubate for 10 minutes.
- Add 100 μL Stop Solution and measure absorbance at 450 nm.
Storage & Notes
- Store all reagents as per instructions in the kit manual.
- Handle Stop Solution with caution as it is corrosive.
- Ensure pH of the sample is between 7.0-7.4 for optimal performance.
- Unused strips should be stored in a resealable bag with desiccant.
