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ProteanFect™ CRISPRMax Mouse Immunocyte Gene Editing Transfection Kit
ProteanFect™ CRISPRMax Mouse Immunocyte Gene Editing Transfection Kit offers a non-viral, non-electroporation, and non-liposomal transfection system utilizing engineered mammalian proteins. This innovative design achieves high transfection efficiency while maintaining a superior safety profile. The ProteanFectTM CRISPRMax-Cas9 mRNA Mouse Immunocyte Gene Editing Kit is specifically designed for effective gene editing by delivering Cas9 mRNA and sgRNA into primary mouse immune cells. It excels in engineering sensitive mouse immune cells, including T cells, NK cells, and γδ T cells.
| Number | Standard | Price(USD) | Stock |
|---|---|---|---|
| PT06-0010B | 0.1 mL | 860 | in two weeks |
| PT06-0020B | 0.2 mL | 1663 | in two weeks |
| PT06-0050B | 0.5 mL | 3189 | in two weeks |
| PT06-0100B | 1 mL | 5077 | in two weeks |
| PT06-0500B | 5 mL | 22200 | in two weeks |
Transfection Performance
Documents
Product Information
Components & Storage
The kit is shipped on dry ice. Once received, store the components as indicated below. The kit includes positive control samples with EGFP-encoding mRNA to verify transfection efficiency as well as single-guide RNA (sgRNA) targeting the mouse Trac gene.
| Components | Storage |
|---|---|
| Reagent A (for Mouse Immunocyte CRISPR-Cas9 mRNA) | 2-8℃ |
| Reagent B (for Mouse Immunocyte CRISPR-Cas9 mRNA) | -20°C(Avoid repeated freeze-thaw cycles |
| Reagent C (for Mouse Immunocyte CRISPR-Cas9 mRNA) | 2-8°C |
| EGFP mRNA (1 µg/µL) | -80°C (Avoid repeated freeze-thaw cycles) |
| Mouse Trac-sgRNA (1 µg / µL) | -80°C (Avoid repeated freeze-thaw cycles) |
Shipping Information
Shipped on Dry Ice
Compatible Cell Lines
Cell Lines Compatible with ProteanFect™ CRISPRMax Mouse Immunocyte Gene Editing Transfection Kit
| Cell lines | Tested Nucleic Acid Types for Transfection |
|---|---|
| Mouse primary T cells | sgRNA |
| Mouse primary NK cells | sgRNA |
| Mouse primary γδ T cells | sgRNA |
| Mouse primary Treg cells | sgRNA |
| Mouse primary CD4+ cells | sgRNA |
| Mouse primary CD8+ cells | sgRNA |
Product Data
Transfection Protocol for mRNA per Well of a 96-Well Plate
| Scheme | Step | Transfection Protocol for ProteanFect™ CRISPRMax Mouse Immunocyte Gene Editing Transfection Kit in Primary mouse immunocyte per Well of a 96-Well Plate |
|---|---|---|
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1. Transfection Complex Preparation |
1.1 Mix Reagent A (for Mouse Immunocyte CRISPR-Cas9 mRNA) with mRNA Mix 0.25 µg Cas9 mRNA and 0.25 µg sgRNA with 40 µL of Reagent A (for Mouse Immunocyte CRISPR-Cas9 mRNA). Note: Invert Reagent A (for Mouse Immunocyte CRISPR-Cas9 mRNA) briefly before use to ensure uniformity. 1.2 Add Reagent B (for Mouse Immunocyte CRISPR-Cas9 mRNA) Add 0.7 μL of Reagent B (for Mouse Immunocyte CRISPR-Cas9 mRNA) to the mixture. Mix gently by pipetting up and down 20-30 times or vortexing for 10 seconds. 1.3 Add Reagent C (for Mouse Immunocyte CRISPR-Cas9 mRNA)Add 10 μL of Reagent C (for Mouse Immunocyte CRISPR-Cas9 mRNA) to the mixture. Mix gently by pipetting up and down 2-3 times or vortexing for 2-3 seconds. Note: If precipitation occurs in Reagent C (for Mouse Immunocyte CRISPR-Cas9 mRNA), heat to 65°C until fully dissolved before use. |
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2. Cell Preparation |
2.1 Suspension cells Harvest the cells by centrifugation at 300 g for 5 minutes. Discard the supernatant and wash the cells once with Opti-MEM. Resuspend the cells with Opti-MEM and adjust the cell concentration to 1×107 – 1.5×10⁷ cells/mL. Note: Avoid including FBS in the transfection medium. 2.2 Adherent cells Maintain 50%-80% cell confluence. Remove medium, wash the cells once with Opti-MEM, and then add 20 µL of Opti-MEM. Note: Avoid including FBS in the transfection medium. Optional: Harvest the cells by trypsinization, then resuspend them in Opti-MEM at a concentration of 1×107 – 1.5×10⁷ cells/mL for subsequent transfection. |
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3. Transfection |
3.1 Mix transfection complex with cells For suspension cells, mix 40 µL of transfection complex with 20 µL of cell suspension and gently pipet up and down 2-3 times. For adherent cells, apply it directly onto the cells. 3.2 Incubation Incubate the cells with the transfection complex for 15-30 minutes in a cell culture incubator. 3.3 Termination Terminate the reaction by adding ≥200 µL of culture medium (10X cell suspension), centrifuge at 300 g for 5 minutes, and discard the supernatant. For adherent cells, replace the transfection mixture with ≥200 µL of culture medium (10X cell suspension). Note: The cell pellet may adhere to the tube walls. Gently remove the supernatant to minimize cell loss. 3.4 Post-transfection culture Incubate the transfected cells in culture medium and evaluate the editing efficiency of the target genes after 48 to 72 hours, or at an appropriate time. |
| Scheme | Step |
|---|
Technical FAQ
1.1 Optimize Transfection Parameters
Optimize transfection parameters for each cell type. Extended incubation time: Adjust the incubation time of the transfection complex with cells. The maximum incubation time is 30 minutes for primary cells. Increase ProteanFect transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Severe Cytotoxicity Caused by Plasmid DNA
The transfection of pDNA into primary cells, such as primary T cells, can induce cytotoxicity and inflammatory responses. Due to the risk of significant toxicity, pDNA transfection is generally not recommended for primary T cells.
1.3 Improve Cell Condition
For primary mouse immunocytes, proper activation is crucial for optimal transfection efficiency. For example, mouse primary T cells generally achieve the best transfection results after stimulation with anti-CD3/CD28 activation beads or antibodies, with peak efficiency typically observed around days 2-4.
1.4 Use Positive Control
We recommend using a 96-well plate format to optimize transfection conditions for a specific cell type, with EGFP mRNA as the positive control.
Optimize transfection parameters for each cell type. Extended incubation time: Adjust the incubation time of the transfection complex with cells. The maximum incubation time is 30 minutes for primary cells. Increase ProteanFect transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Severe Cytotoxicity Caused by Plasmid DNA
The transfection of pDNA into primary cells, such as primary T cells, can induce cytotoxicity and inflammatory responses. Due to the risk of significant toxicity, pDNA transfection is generally not recommended for primary T cells.
1.3 Improve Cell Condition
For primary mouse immunocytes, proper activation is crucial for optimal transfection efficiency. For example, mouse primary T cells generally achieve the best transfection results after stimulation with anti-CD3/CD28 activation beads or antibodies, with peak efficiency typically observed around days 2-4.
1.4 Use Positive Control
We recommend using a 96-well plate format to optimize transfection conditions for a specific cell type, with EGFP mRNA as the positive control.
Transfected cells may exhibit transient changes in behavior, but typically, viability will be restored by the second day post-transfection.
In 96-well formats, it is common for the pellet to be less distinct and may adhere to the tube walls. Gently pipetting can help minimize cell loss.
For further questions, please contact us at: tech@nanoportalbio.com.