Use case
Use A549 to test compound/genetic effects on proliferation/survival.
In one line
Expand → seed → dose matrix → viability/apoptosis.
Protocol overview
- 01Expand; log passage
- 02Seed overnight
- 03Dose + vehicle
- 04Incubate 24–72 h
- 05Viability assay
- 06Fit IC₅₀
- 07Independent repeats
- 01[Prep] Confirm A549 medium; mycoplasma-negative.
- 02[Seed] Plate to ~40–70% confluence at dosing; overnight attach.
- 03[Dose] Vehicle + ≥5-point log grid; DMSO ≤0.1–0.5% if possible.
- 04[Incubate] Typically 24–72 h.
- 05[Viability] MTT/CCK-8/CTG; ≥3 wells/group.
- 06[Optional] Apoptosis or colony formation.
- 07[Calculate] % inhibition / IC₅₀; ≥2–3 repeats.
- 08[QC] Log passage and serum lot.
Reagents / compounds
| Name | Role | Conc. |
|---|---|---|
| Test compound | Drug/mechanism | 对数梯度,预实验优化 |
| Vehicle | Vehicle control | 等体积分数 |
| Viability reagent | Quantify viability | 按说明书 |
Readouts
Viability, IC₅₀, optional apoptosis/colonies.
Literature & manuals
- [1]PubMedPMID 437016Int J Cancer · 1976A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells.
- [2]PubMedPMID 2141510In Vitro · 1980Growth of a human carcinoma cell line in serum-free medium.
- [3]PubMedPMID 6606682J Immunol Methods · 1983Rapid colorimetric assay for cellular growth and survival.