Low Range Cas9 ELISA Assay
The Low Range Cas9 ELISA Assay is an ultra-sensitive enzyme-linked immunosorbent assay (ELISA) specifically designed for the detection and quantification of low levels of S. pyogenes Cas9 protein in cell culture lysates. This assay is ideal for researchers who require precise measurement of minimal Cas9 expression, making it a valuable tool for CRISPR/Cas9 gene editing studies and experimental screening.
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 is specifically captured onto the immobilized antibody. After an incubation period, unbound components are washed away.
A biotinylated mouse monoclonal Cas9 detection antibody is then added, followed by a Streptavidin-HRP conjugate, which binds to the biotinylated antibody. The addition of tetramethylbenzidine (TMB) substrate produces a colorimetric reaction proportional to the Cas9 concentration in the sample. The absorbance is measured spectrophotometrically, providing quantitative detection of low Cas9 levels with high accuracy.
Applications:
✔ Detection of Low Cas9 Expression in Cell Culture Lysates
✔ Monitoring CRISPR/Cas9 Gene Editing Experiments
✔ Screening Experimental Samples for Minimal Cas9 Presence
✔ Preclinical and Laboratory Research in Genome Editing
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.