This kit contains recombinant Mouse IL-4, recombinant Mouse IFN-γ, and LPS. It is designed to efficiently and specifically induce mouse RAW264.7 cells in the M0 state (unpolarized, resting) into classical M1-type (pro-inflammatory) and M2-type (anti-inflammatory/reparative) macrophages in vitro.

IFN-γ combined with LPS is used to polarize RAW264.7 cells into M1 macrophages. IFN-γ serves as the "priming" signal, upregulating the macrophages' responsiveness to subsequent stimuli. LPS acts as the "trigger" signal, initiating a robust pro-inflammatory response via the TLR4 pathway.

IL-4 is used to polarize RAW264.7 cells into M2 macrophages. IL-4 drives the differentiation of macrophages towards the alternatively activated M2 phenotype by activating the STAT6 pathway.

RAW264.7 cells are easy to culture, passage, and transfect, and exhibit clear responses to stimulation signals, making them a gold standard model for studying macrophage biology, immune responses, inflammatory diseases, cancer immunity, and therapeutic screening. Using this kit streamlines experimental workflows and ensures comparability of results across different experimental batches.

Product Information

Components

Product Includes

Reconstitution Method

Storage

Notes

1. For your safety and health, please wear a lab coat and disposable gloves during operation.

2. This product is for research use only.

Instructions

Induction of RAW264.7 Cells into Macrophages

I. M1 Polarization

1. Seeding: Seed cells in a 6-well plate at a density of 2 × 10⁵ cells/well. Incubate at 37°C with 5% CO₂ for 24 hours to allow full attachment.

Note: Do not overload the cells; excessive cell density may affect polarization efficiency.

2. M1 Induction: Replace the medium with DMEM + 2% FBS (to minimize the impact of cytokines in the serum). Add recombinant Mouse IFN-γ (25 ng/mL) and LPS (100 ng/mL).

Control: Set up a control group (cells only, without inducers).

Incubate at 37°C with 5% CO₂ for 24 hours.

II. M2 Polarization

1. Seeding: Seed cells in a 6-well plate at a density of 2 × 10⁵ cells/well. Incubate at 37°C with 5% CO₂ for 24 hours to allow full attachment.

Note: Do not overload the cells; excessive cell density may affect polarization efficiency.

2. M2 Induction: Replace the medium with DMEM + 2% FBS (to minimize the impact of cytokines in the serum). Add recombinant Mouse IL-4 (40 ng/mL).

Control: Set up a control group (cells only, without inducers).

Incubate at 37°C with 5% CO₂ for 24 hours.

Documents:

Manuals:

92648_Manual_HB20260409

Related Blogs:

M1 or M2 Macrophages? The Complete RAW264.7 Polarization Solution Researchers Need