The Standard Inflammasome Antibodies for Western Blotting

Inflammasomes and Caspase-1 Activation

Inflammasomes are large molecular complexes that activate inflammatory caspases and cytokines of the IL-1 family. Key inflammasomes include NLRP1, NLRP3, IPAF, and AIM2, each defined by their specific protein components. These complexes are activated by various signals such as bacteria, toxins, and other danger signals, leading to the recruitment of adapter proteins like ASC and the activation of caspase-1, a dominant inflammatory caspase.

Activity Regulation and Therapeutic Implications

The regulation of inflammasome activity involves various proteins that can interfere with inflammasome activation and caspase processing. Understanding these mechanisms is vital, as inflammasomes are implicated in several diseases, including cryopyrin-associated periodic syndromes (CAPS), gout, and Type 2 Diabetes Mellitus (T2DM). Targeting inflammasome components offers potential therapeutic strategies for these conditions.

Measuring Inflammasomes with Standard Antibodies

Detecting inflammasome activation, particularly through caspase-1 immunoblotting, is a standard yet technically challenging method. AdipoGen Life Sciences provides KO-extract validated antibodies, ensuring specificity and reliability in Western blotting applications. Their NLRP3 Inflammasome Starter Sets for both human (AG-44B-0008) and mouse (AG-44B-0009) models offer comprehensive solutions for studying inflammasomes, including key components like NLRP3, ASC, and caspase-1. Additionally, the NLRP3 Inflammasome Human Reagents Starter Set (AG-44B-0010) and Mouse Reagents Starter Set (AG-44B-0011) include chemical compounds for priming and activation.

KO-Extract Validated Antibodies

AdipoGen Life Sciences offers a range of KO-extract validated antibodies specifically designed for inflammasome research:

• Anti-Caspase-1 (p20) (mouse), mAb (Casper-1)
• Anti-Caspase-1 (p10) (mouse), mAb (Casper-2)
• Anti-Caspase-1 (p20) (human), mAb (Bally-1)
• Anti-NLRP3/NALP3, mAb (Cryo-2)
• Anti-Asc, pAb (AL177)

Tips for Successful Western Blotting

• Sample Preparation: Ensure cells or tissues express detectable levels of NLRP3. Priming with agents like LPS or PMA can enhance expression.
• SDS-PAGE and Transfer: Use gels with less than 8% SDS-PAGE concentration and perform transfers overnight at 4°C for optimal results.
• Antibody Usage: Utilize freshly prepared antibodies at recommended dilutions to ensure specificity and sensitivity.
• Positive Controls: Employ cell lines known for high NLRP3 expression, such as THP1 or Raw264.7, to validate experiments.

AdipoGen Life Sciences’ antibodies are among the few validated using KO extracts, providing researchers with reliable tools for inflammasome research. Their products, including ready-to-use LPS solutions and TNF-α proteins, further support inflammasome priming and activation studies.

By leveraging these standard antibodies and protocols, researchers can advance their understanding of inflammasomes and their roles in health and disease, paving the way for novel therapeutic interventions