Cas9 ELISA Assay
The Cas9 ELISA Assay is a highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) designed for the quantification of S. pyogenes Cas9 protein in cell culture lysates. This assay provides a reliable and accurate method for monitoring Cas9 expression, making it an essential tool for researchers working with CRISPR/Cas9 gene editing systems.
Assay Principle:
This ELISA utilizes murine anti-Cas9 capture antibodies pre-coated onto microtitration wells. When test samples containing Cas9 antigen are introduced, the Cas9 protein specifically binds to the immobilized antibody. After an incubation period, unbound components are washed away.
A biotinylated mouse monoclonal Cas9 detection antibody is then added, enhancing specificity in antigen detection. Following a wash cycle, a Streptavidin-HRP conjugate is introduced, which binds to the biotinylated antibody. The addition of tetramethylbenzidine (TMB) substrate generates a colorimetric reaction proportional to the Cas9 concentration in the sample. The absorbance is measured spectrophotometrically, providing a quantitative readout of Cas9 protein levels.
Applications:
✔ Detection and Quantification of S. pyogenes Cas9 in Cell Lysates
✔ Screening Experimental Samples Utilizing CRISPR/Cas9 Gene Editing
✔ Optimization of Cas9 Expression for Genome Editing Studies
✔ Preclinical and Laboratory Research in Gene Editing Technologies
Kit Presentation
Materials Supplied: The reagents supplied in this pack are for research use only.
1-Coated microwell strips - 1 plate
plastic microtitration wells coated with anti-Cas9 murine monoclonal antibody in foil pouch with desiccant - 96 wells
2-Cas9 Positive Control - 2.5 µg/mL - 0.1 mL
3-Lysis Buffer - (NaCl, NP-40 Alt., NaDeoxycholate, SDS, Tris-HCl, pH 7.5) - 30 mL
4-Detector - anti-Cas9 murine monoclonal antibody conjugated to biotin - 12 mL
5-Conjugate - streptavidin conjugated to horseradish peroxidase enzyme containing 0.01% Bromonitrodioxane as preservative - 12 mL
6-Wash Buffer (20x concentrated) - Tris buffered saline containing 0.05% Tween-20, must be diluted before use - 60 mL
7- Substrate Solution - Ready to use, Tetramethylbenzidine - 12 mL
8-Stop Solution - 1 N H2SO4 - 12 mL
Storage and Stability
The Cas9 Positive Control can be stored at +4˚C for 1 month. For long term storage, immediately store at -2˚C or colder. The Cas9 Positive Control is stable for a maximum of 3 freeze-thaw cycles. If more cycles are required, aliquot into smaller portions and freeze tubes at -20ºC or colder until needed. All other reagents should be stored at 2-8˚C and should not be used beyond the expiration date on the label. Once opened, microtitration strips may be stored at 2-8˚C until the expiration date on the label, provided that desiccated conditions are maintained. Unused strips should be returned to their original foil pouch along with the sachet of desiccant. Opened pouches should be securely resealed. The 1X Wash Buffer should not be stored for longer than 3 weeks at 2-8˚C. It is recommended that Wash Buffer be freshly diluted before each assay. Do not use If the 1X Wash Buffer becomes visibly cloudy or develops precipitate during the 3 weeks.