Cas9 Nuclease (10mg/mL)-14701ES

Cas9 Nuclease, derived from wild-type Streptococcus pyogenes, is an RNA-guided endonuclease capable of specifically cleaving double-stranded DNA (it can also cleave single-stranded DNA or RNA in the presence of DNA PAM). The Cas9 cleavage site is located within the target sequence, three bases away from the PAM (NGG) region. This product, through codon optimization and nuclear localization signal (NLS) design, is recombinantly expressed in Escherichia coli, exhibiting high editing efficiency. It can be used for gene modification in cells (such as hematopoietic stem cells, T cells, etc.), as well as in molecular diagnostics and pathogen detection.

Product Features:

• Optimized Core Positioning Signal: Enhanced NLS (Nuclear Localization Signal) for improved editing efficiency.
• Consistent Editing Efficiency: Consistent high editing efficiency both in vitro and in vivo.
• His Tag: His tag provides flexibility in fusion protein detection methods.
• High Purity: Purity exceeding 95%.
• High Concentration: Suitable for standard editing conditions and adaptable for optimizing editing conditions in challenging scenarios such as primary or embryonic cells, microinjection, or screening multiple gRNA sequences simultaneously.

Product Information

Specifications	100 μg / 500 μg / 1 mg
Source	Recombinant expression in Escherichia coli of Cas9 gene from Streptococcus pyogenes
Enzyme Storage Buffer	30 mM Tris-HCl, 300 mM NaCl, 0.1 mM EDTA, 50% Glycerol, pH 7.4
Concentration	10 mg/mL
Endotoxin	≤10 EU/mg
Purity	≥95%
Tag	His

 

Inquiry of GMP-Grade Cas9

Component Information

Component Name	14701ES60	14701ES76	14701ES03
Cas9 Nuclease (10 mg/mL)	10 μL	50 μL	100 μL

 

Product Applications

• Genome editing based on CRISPR/Cas9 technology
• Gene modification of cells and gene therapy drugs (hematopoietic stem cells, T cells, etc.)
• In vitro screening of effective sgRNA
• In vitro cleavage of target DNA

Storage Conditions

Store at -25 to -15°C with a shelf life of 1 year.

Instructions for Use

RNP Preparation:

1. Dissolve sgRNA powder in 1×TE Buffer (pH 7.5) to a final concentration of 100 μM, thoroughly vortex.
2. Prepare the following system and mix well:

Components	Volume (μL)
Cas9 Nuclease (10 mg/mL)	1.28
sgRNA (100 μM)	2.34
PBS	1.38
Total	5

• Note: The molar ratio of Cas9 Nuclease to sgRNA is approximately 1:3.
• Incubate at room temperature for 20 minutes.

Precautions

1. To prevent RNase contamination, maintain a clean experimental area, wear clean gloves and masks during operation, and use RNase-free consumables such as pipette tips and centrifuge tubes.
2. Avoid repeated freeze-thaw cycles. After initial dissolution, it is recommended to aliquot and store according to usage.
3. For your safety and health, wear lab coats and disposable gloves during operation.
4. This product is for research purposes only.