ImmunoFect STAR™ Intracellular Antibody Delivery
This is intended as a guide only; for full experimental details please read the reference provided.
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Intracellular Antibody Delivery with ImmunoFect STAR™
Background
ImmunoFect STAR™ is a highly efficient antibody transfection reagent designed for intracellular delivery of antibodies into live cells. ImmunoFect STAR™ enables functional studies of intracellular targets by facilitating direct antibody access to the cytosol. The delivery mechanism is optimized for high efficiency while maintaining cell viability.
ImmunoFect STAR ™ / Antibody Complex Formation
ImmunoFect STAR™ is supplied in anhydrous ethanol, 10 mg/mL.
- Dilute with HEPES buffer (20 mM, pH 7.4), (5 μL ImmunoFect STAR™ to 45 μL HEPES buffer), to make a 1 mg/mL ImmunoFect STAR™ solution.
- Prepare the ImmunoFect STAR™ antibody complex by diluting 3 μL of 1 mg/mL ImmunoFect STAR™ with 27 μL HEPES buffer. Combine the 30 μL of diluted ImmunoFect STAR™ solution with 20 μL of antibody solution (2 μL of 1 mg/mL antibody, with 18 μL HEPES buffer) and incubate for 1 minute.
- Add ImmunoFect STAR™ antibody complex solution to 450 μL serum-free DMEM or Leibovitz’s L-15 medium and apply to cells.
It may be necessary to optimize the ratio (ImmunoFect STAR™ concentration of 4-8 μg/mL, and antibody concentration of 2-8 μg/mL) to achieve maximum efficiency for intracellular delivery.