NEGES0890 Protein A (PA) ELISA Kit
The Protein A ELISA kit is applicable in the field of biotechnology and biomedicine, it is for quantitative measurement of Protein A residues in the purification process of biopharmaceutical production, for examples like purified fermentation broth and cell culture supernatant, etc. This ELISA kit is for research use only, it should not be used in clinical diagnostic procedures.
Product Information | |
Cat. NO. | NEGEP0890 |
Product Name | Protein A (PA) ELISA Kit |
Species | General |
Product Size | 96 Tests |
Concentration | 0.1875-6ng/ml |
Sensitivity | 47pg/ml |
Principal | Sandwich ELISA |
Sample Volume | 100 ul |
Sample Type | ELISA Kit for the quantitative Measurement of Protein A Residues in Protein Purification Process, and End-Product (purified fermentation broth, cell culture supernatant, etc.) |
Assay Time | 3 hours |
Platform | Microplate Reader |
Conjugate | HRP |
Detection Method | Colorimetric |
Storage | 2-8°C |
Kit Components | ||
Reagents | Specification | Quantity |
Pre-Coated Microplate (Detachable) | 96 wells | 1 plate (Keep Sealed) |
| Standard (Stock Solution) -2ug/ml | 20ul | 1 vial |
Standard S1 | 6ng/ml | 1 vial |
Standard S2 | 3ng/ml | 1 vial |
Standard S3 | 1.5ng/ml | 1 vial |
Standard S4 | 0.75ng/ml | 1 vial |
Standard S5 | 0.375ng/ml | 1 vial |
Standard S6 | 0.1875ng/ml | 1 vial |
HRP-conjugated antibody(400×) | 50ul | 1 vial |
| Sample Pretreatment Solution | 10ml | 1 vial |
TMB Substrates | 10ml | 1 vial (Avoid Light) |
Stop Solution | 10ml | 1 vial |
Wash Solution (100×) | 10ml | 1 vial |
| Diluent Buffer(10x) | 10ml | 1 vial |
Plate Sealer | 4 pieces | |
Instruction Manual | 1 | |
Principle of the Assay |
This ELISA kit is applied to the double antibody sandwich Enzyme Linked Immunosorbent Assay to detect the concentration of protein A in samples. Before starting the ELISA experiment, mix the samples and serially diluted standards with denaturing solution, and incubate at room temperature to completely dissociate Protein A from the antibodies. After pretreatment, add the detection antibody to the microplate wells pre-coated with chicken anti-Protein A antibody. Then add the denatured standards and samples. Incubate at room temperature to allow the specific binding of antigen-antibody, capturing Protein A from the samples and standards onto the microplate. After incubation, a sandwich complex forms with the coating antibody, Protein A, and detection antibody. Wash the plate to remove unbound substances. Then a TMB substrate solution is added to the wells to incubate. TMB substrate solution turns blue after the oxidation of HRP, and finally turns to yellow immediately after adding the stop solution. Color of TMB substrate positively correlated with total protein A bound in the initial steps. The color is measured by spectrophotometrically with wavelength of 450nm. The concentration of protein A in samples is then determined by comparing the O.D. of the samples to the standard curve. |
Coefficient of Variance | Intra Variation: 7-8% | ||
Inter Variation: 6-9% | |||
Recovery | In the first experiments, the matrix effect on experimental results should be confirmed through spike recovery. It is recommended to add one volume of standard solution (6 ng/mL) to every three volumes of the sample. Calculate the recovery rate using the formula: (actual concentration after spiking - concentration before spiking) / theoretical concentration (3 ng/mL). A recovery rate of 80-120% is considered acceptable. | ||
Specificity/Cross-reactivity | Capture antibody and detection antibody respectively identify different epitopes of the antigen, which maximizes the specificity of the reaction. | ||
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