Protein A ELISA Kit
Staphylococcus aureus Protein A ELISA Kit Colorimetric assay for quantification of Staphylococcus aureus Protein A.
Catalog No. ABIN2044912
Quick Overview for Protein A ELISA Kit (ABIN2044912)
Target
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Protein A
Reactivity
Staphylococcus aureus
Detection Method
Colorimetric
Method Type
Sandwich ELISA
Application
ELISA
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Analytical Method
- Quantitative
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Components
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Anti-Protein A Microwell Strip Plate, 8x12 strips
Protein A Stock, lyophilized
Anti-Protein A HRP Conjugate
Sample Diluent (20X)
Wash Solution (100X)
TMB Substrate
Sample Recovery:
Protein A immunosorbents are widely used for purification of monoclonal and polyclonal antibodies. Possible leakage of Protein A during processing may contaminate the purified antibody preparations. The detection/measurement of contaminating Protein A is complicated by Protein A-IgG binding in the sample, which inhibits the subsequent binding of the Protein A to the ELISA antibodies required for accurate quantification. A sample pretreatment boiling step for Protein A samples containing mouse IgG heat denatures the IgG. This uncouples the Pro A-IgG binding and, thus, allows for accurate Protein A measurement. Appropriate recovery of Protein A in solutions of IgG from other species has not been elucidated with this ELISA. In these cases, the investigator should demonstrate requirements for appropriate Protein A recovery.
High and low concentrations of Protein A were mixed into buffer containing mouse IgG at 1mg/ml. Samples were assayed before and after a boiling pretreatment step. Observed assay values in samples containing IgG compared to non-IgG sample values ranged from 41 to 62%. With a boiling pretreatment step, observed IgG sample assay values were 75 to 106% of non-IgG values, indicating accurate quantification of Protein A in the presence of IgG using the pretreatment step.
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Protocol
- The Protein A ELISA Kit is a Sandwich ELISA for the highly sensitive detection and/or quantification of Protein A in fluids. The kit is designed to measure trace contamination of Protein A in antibody solutions. The antibodies used in this kit were produced by immunization with recombinant Protein A. Native and recombinant Protein A are quantified in this assay with similar potency.The Protein A ELISA kit is based on the binding of Protein A in samples to two antibodies, one immobilized on the microtiter wells, and the other conjugated to horseradish peroxidase (HRP) enzyme. After a washing step, chromogenic substrate is added and color is developed by the enzymatic reaction of HRP on the TMB substrate, which is directly proportional to the amount of Protein A present in the sample. Stopping Solution is added to terminate the reaction, and absorbance at 450nm is then measured using an ELISA microtiter well reader. The concentration of Protein A in samples is calculated from a curve of standards containing known concentrations of Protein A.
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Restrictions
- For Research Use only
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Storage
- 4 °C/-20 °C
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- Protein A
Target See all Protein A products
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