尼日利亞菌素

Nigericin 是一種有效的微生物毒素，可作為鉀離子載體，誘導細胞內鉀水平的淨降低。這對於 NLRP3 炎性體的寡聚化和 caspase-1 在焦亡中的激活至關重要。Nigericin 可用作細胞焦亡實驗中的陽性對照。

Background

Inflammatory caspases, such as caspase-1, play a critical role in the mechanism to trigger the lytic programmed cell death known as pyroptosis. Nigericin can be utilized as a positive control in pyroptosis experiments. Nigericin is a potent microbial toxin derived from Streptomyces hygroscopicus. It acts as a potassium ionophore, inducing a net decrease in intracellular levels of potassium, which is crucial for oligomerization of the NLRP3 inflammasome and activation of caspase-1. It requires signaling through pannexin-1 to induce caspase-1 activation and IL-1ß processing and release. It has been shown to generate a robust caspase-1 activation response in various cell lines, including Jurkat and THP-I cells.

Reagent Name

Nigericin

Storage

-20°C

Country of Origin

United States

Protocols

1. Reconstitute each vial of Nigericin with 100 µL DMSO to form the 5 mM stock concentrate. Once reconstituted, it may be aliquoted and stored at ≤ -20°C for 1 year protected from light and thawed no more than twice during that time.

2. Immediately prior to addition to the samples and controls, dilute 5 mM Nigericin stock 1:10 in diH2O to form a 500 µM working solution for use in treating samples. For example, dilute 1:10 by adding 20 µL stock concentrate to 180 µL diH2O.

3. Use Nigericin at 1-20 µM to induce NLRP3 inflammasome activation in cells. For example, to use at 10 µM, dilute 500 µM working solution 1:50 in samples; e.g., spike 294 µL cell suspension/overlay medium with 6 µL of 500 µM working solution. Typical treatment periods range from 3-24 hours at 37°C. Each investigator should adjust the concentration of Nigericin and treatment period to accommodate the particular cell line and research conditions.

4. NLRP3 inflammasome activation can be detected by ELISA or Western blot measuring secreted pro-inflammatory cytokines IL-1β or IL-18, or through the use of caspase-1 activation assays, such as ICT’s Caspase-1 Assay Kits or Pyroptosis/Caspase-1 Assay Kit.