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ProteanFect™ CRISPRMax Gene Editing Transfection Kit
ProteanFect™ CRISPRMax 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 Gene Editing Kit is specifically designed for effective gene editing by delivering Cas9 mRNA and sgRNA into primary cells and difficult-to-transfect cell lines. Additionally, it excels in co-transfecting sgRNAs targeting different genes, enabling simultaneous knockout of multiple genes.
| Number | Standard | Price(USD) | Stock |
|---|---|---|---|
| PT05-0010B | 0.1 mL | 842 | in two weeks |
| PT05-0020B | 0.2 mL | 1500 | in two weeks |
| PT05-0050B | 0.5 mL | 2835 | in two weeks |
| PT05-0100B | 1 mL | 4483 | in two weeks |
| PT05-0500B | 5 mL | 19648 | 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 human TRAC gene.
| Components | Storage |
|---|---|
| Reagent A (for CRISPR-Cas9 mRNA) | 2-8℃ |
| Reagent B (for CRISPR-Cas9 mRNA) | -20°C(Avoid repeated freeze-thaw cycles |
| Reagent C (for CRISPR-Cas9 mRNA) | 2-8°C |
| EGFP mRNA (1 µg/µL) | -80°C (Avoid repeated freeze-thaw cycles) |
| Human TRAC-sgRNA (1 µg / µL) | -80°C (Avoid repeated freeze-thaw cycles) |
Shipping Information
Shipped on Dry Ice
Compatible Cell Lines
Primary Cells and Cell Lines Compatible with ProteanFect™ CRISPRMax Gene Editing Transfection Kit
| Cell lines | Tested Nucleic Acid Types for Transfection |
|---|---|
| Human primary T cells | sgRNA |
| Human primary NK cells | sgRNA |
| Human primary monocytes | sgRNA |
| Human primary CD34+ hematopoietic stem cells | sgRNA |
| Mouse primary cardiomyocytes | sgRNA |
| Mouse primary neurons | sgRNA |
| Mouse primary glial cells | sgRNA |
| Mouse primary T cells | sgRNA |
| Mouse primary NK cells | sgRNA |
| Large yellow croaker primary mesenchymal stem cells | sgRNA |
| Jurkat T (human T lymphoblastic leukemia cells) | sgRNA |
| LX-2 (human hepatic stellate cells) | sgRNA |
| HepG2 (human liver tumor cells) | sgRNA |
| THP-1 (human acute monocytic leukemia cell line) | sgRNA |
| Raji (human Burkitt lymphoma cells) | sgRNA |
| K562 (human chronic myeloid leukemia cells) | sgRNA |
| MOLT-16 (human T lymphoblastic leukemia cells) | sgRNA |
| SH-SY5Y (human neuroblastoma cells) | sgRNA |
| U2OS (human osteosarcoma cells) | sgRNA |
| U937 (human lymphoma cell line) | sgRNA |
| HFF (human foreskin fibroblasts) | sgRNA |
| HEK-293 (human embryonic kidney cell line) | sgRNA |
| MC38 (mouse colon cancer cells) | sgRNA |
| RAW264.7 (mouse mononuclear macrophage leukemia cells) | sgRNA |
| LLC (mouse Lewis lung cancer cells) | sgRNA |
| C2C12 (mouse myoblasts) | sgRNA |
| COS7 (African green monkey kidney fibroblast-like cells) | sgRNA |
Product Data
Transfection Protocol for mRNA per Well of a 96-Well Plate
| Scheme | Step | Transfection Protocol for ProteanFect™ CRISPRMax Gene Editing in Various Cell Lines per Well of a 96-Well Plate |
|---|---|---|
 |
1. Transfection Complex Preparation |
1.1 Mix Reagent A (for CRISPR-Cas9 mRNA) with mRNA Mix 0.25 µg Cas9 mRNA and 0.25 µg sgRNA with 40 µL of Reagent A (for CRISPR-Cas9 mRNA). Note: Invert Reagent A (for CRISPR-Cas9 mRNA) briefly before use to ensure uniformity. 1.2 Add Reagent B (for CRISPR-Cas9 mRNA) Add 1.4 µL of Reagent B (for CRISPR-Cas9 mRNA) to the mixture. Mix gently by pipetting up and down 20-30 times or vortexing for 10 seconds. |
 |
2. Cell Preparation |
2.1 Suspension cells Harvest the cells by centrifugation at 300 g for 5 minutes. Discard the supernatant and wash cells once with Opti-MEM. Resuspend cells with Opti-MEM and adjust concentration to 5×10⁶ - 1×10⁷ cells/mL. Note: Avoid including FBS in the transfection medium. 2.2 Adherent cells Maintain 50%-80% cell confluence. Remove medium, wash cells once with Opti-MEM, then add 20 µL of Opti-MEM. Note: Avoid including FBS in the transfection medium. Optional: Harvest cells by trypsinization, then resuspend them in Opti-MEM at a concentration of 5×10⁶ - 1×10⁷ cells/mL for subsequent transfection. |
 |
3. Transfection |
3.1 Mix 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 directly to the cells. 3.2 Incubation Incubate the cells with the transfection complex for 45-60 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 cultureIncubate 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 | Transfection Protocol for ProteanFect™ CRISPRMax Gene Editing in Various Primary Cells per Well of a 96-Well Plate |
|---|---|---|
 |
1. Transfection Complex Preparation |
1.1 Mix Reagent A (for CRISPR-Cas9 mRNA) with mRNA Mix 0.25 µg Cas9 mRNA and 0.25 µg sgRNA with 40 µL of Reagent A (for CRISPR-Cas9 mRNA). Note: Invert Reagent A (for CRISPR-Cas9 mRNA) briefly before use to ensure uniformity. 1.2 Add Reagent B (for CRISPR-Cas9 mRNA) Add 0.7 µL of Reagent B (for 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 CRISPR-Cas9 mRNA) Add 8 µL of Reagent C (for CRISPR-Cas9 mRNA) to the mixture. Mix gently by pipetting up and down 2-3 times or vortexing for 2-3 seconds. |
 |
2. Cell Preparation |
2.1 Suspension cells Harvest the cells by centrifugation at 300 g for 5 minutes. Discard the supernatant and wash cells once with Opti-MEM. Resuspend cells with Opti-MEM and adjust concentration to 5×10⁶ - 1×10⁷ cells/mL. Note: Avoid including FBS in the transfection medium. 2.2 Adherent cells Maintain 50%-80% cell confluence. Remove medium, wash cells once with Opti-MEM, then add 20 µL of Opti-MEM. Note: Avoid including FBS in the transfection medium. Optional: Harvest cells by trypsinization, then resuspend them in Opti-MEM at a concentration of 5×10⁶ - 1×10⁷ cells/mL for subsequent transfection. |
 |
3. Transfection |
3.1 Mix 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 directly to 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 cultureIncubate 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. |
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 2 hours for cell lines, and 30 minutes for primary cells. Increase ProteanFect transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Improve Cell Condition
For cell lines, transfect cells with >90% viability, confirmed by trypan blue exclusion. Avoid using cells beyond 15 passages, and allow 2-3 passages for recently thawed cells to stabilize before transfection.
For primary cells, proper activation is crucial for optimal transfection efficiency. For example, human primary T cells generally achieve the best transfection results after stimulation with anti-CD3/CD28 activation beads or antibodies for 2-10 days, with peak efficiency typically observed around days 4-6.
1.3 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 2 hours for cell lines, and 30 minutes for primary cells. Increase ProteanFect transfection complex: Consider increasing the amount of transfection complex to improve transfection efficiency.
1.2 Improve Cell Condition
For cell lines, transfect cells with >90% viability, confirmed by trypan blue exclusion. Avoid using cells beyond 15 passages, and allow 2-3 passages for recently thawed cells to stabilize before transfection.
For primary cells, proper activation is crucial for optimal transfection efficiency. For example, human primary T cells generally achieve the best transfection results after stimulation with anti-CD3/CD28 activation beads or antibodies for 2-10 days, with peak efficiency typically observed around days 4-6.
1.3 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.
Contact Information: For further questions, please contact us at: tech@nanoportalbio.com.