Control Knockout HEK293T RIPA Lysate is a control RIPA lysate prepared from polyclonal HEK293T cells in which the target gene has been knocked out via the CRISPR system, achieved by the simultaneous expression of Cas9, target gene sgRNA, and puromycin resistance gene. This control lysate does not contain sgRNA. This lysate or GFP Knockout HEK293T RIPA Lysate is commonly used as a control lysate for target gene knockout using CRISPR/Cas9 technology.
This RIPA lysate is derived from polyclonal HEK293T cells infected with a lentivirus capable of co-expressing Cas9 and puromycin resistance gene, followed by puromycin screening. Please refer to the figure below for key map information of the gene sequence of the corresponding lentivirus used to generate these cells.
Key map information of the gene sequence of the lentivirus capable of simultaneously expressing sgRNA, Cas9 and puromycin resistance.
Note: No sgRNA is present in this control lysate.
Instructions for Use:
1. Protein Concentration Determination of Cell RIPA Lysate Samples:
It is recommended to determine the protein concentration using the ready-to-use BCA Protein Quantification Kit (R1491648) produced by Aladdin. Due to the high concentration of detergent in this product, the Bradford method is not recommended for protein concentration determination.
Application of Cell RIPA Lysate:
After protein concentration determination, add SDS-PAGE protein loading buffer to this RIPA lysate. It can then be used for SDS-PAGE, Western blot detection, validation of antibodies against the target gene, etc.
Precautions:
1. This product is for scientific research use only by professional personnel. It is not intended for clinical diagnosis or therapy, nor for use in food or pharmaceutical products. Do not store in ordinary residential premises.
2. For your safety and health, please wear a lab coat and disposable gloves during operation.
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