Human THP-1 Cell Polarization Induction Kit
Human THP-1 Cell Polarization Induction Kit
Product Details
Product Details
Product Specification
| Species | Human |
| Reconstitution | IL-4/IL-13/IFN-γ Protein: Reconstitute at 0.1-1 mg/ml according to the size in ultrapure water after rapid centrifugation. |
Components
Contains |
Reference dosage |
SSpecification |
MSpecification |
LSpecification |
IL-13 Protein, Human |
20 ng/mL |
10μg |
50μg |
100μg |
IL-4 Protein, Human |
20 ng/mL |
10μg |
50μg |
100μg |
IFN-γ Protein, Human |
20 ng/mL |
10μg |
50μg |
100μg |
Lipopolysaccharide (LPS) solution (500X, 2.5mg/ml) |
100 ng/mL |
100μl |
100μl |
100μl*2 |
Protocol
Preparation before polarization:
The passage number of cells used for polarization induction should not be too high. Before inducing polarization, use flow cytometry to detectTHP-1cellCD14andCD11bexpression. OnlyTHP-1cells that do not express or have low expression of both markers can be used for subsequent experiments.
M1/M2Induction
1.M1Induction:
UseM0induction medium (1640complete medium containing100 ng/mL PMA), seedTHP-1>95%,<20passages) at a density of 5×10⁵ cells/mL into10 cm culture dishes (10 mL), culture for24h to generate macrophagesM0 (cells adhere, pseudopodia extend). Gently wash with pre-warmedPBSfor1-2times to removePMA. Replace withM1induction medium (1640complete medium containing100 ng/mL LPS,20 ng/mL IFN-γ), culture for48h.
Negative control: UseM0induction medium (1640complete medium containing100 ng/mL PMA), seedTHP-1>95%,<20passages) at a density of 5×10⁵ cells/mL into10 cm culture dishes (10 mL), culture for24h to generate macrophagesM0 (cells adhere, pseudopodia extend). Gently wash with pre-warmedPBSfor1-2times to removePMA. Replace with complete medium and culture for48h. It is recommended to useAccutasefor sample collection and testing.
2.M2Induction:
UseM0induction medium (1640complete medium containing100 ng/mL PMA), seedTHP-1>95%,<20passages) at a density of 5×10⁵ cells/mL into10 cm culture dishes (10 mL), culture for24h to generate macrophagesM0 (cells adhere, pseudopodia extend). Gently wash with pre-warmedPBSfor1-2times to removePMA, replace with complete medium and rest for24h. Then replace withM2induction medium (1640complete medium containing20 ng/mL IL-4,20 ng/mL IL-13), culture for24h.
Negative control: UseM0induction medium (1640complete medium containing100 ng/mL PMA), seedTHP-1>95%,<20passages) at a density of 5×10⁵ cells/mL into10 cm culture dishes (10 mL), culture for24h to generate macrophagesM0 (cells adhere, pseudopodia extend). Gently wash with pre-warmedPBSfor1-2times to removePMA, replace with complete medium and rest for48h. It is recommended to useAccutasefor sample collection and testing.
