Human Interleukin 1beta converting enzyme, ICE ELISA Kit

The Human Interleukin 1beta Converting Enzyme (ICE) ELISA Kit is a highly sensitive and specific assay designed for the quantitative detection of ICE levels in human serum, plasma, culture media, urine, and other biological fluids. This kit is essential for research into inflammatory responses, apoptosis pathways, and related physiological and pathological processes.

Purpose

• Quantification of ICE Levels:
  • Detect and measure ICE concentrations in human biological samples to study inflammatory signaling, caspase activation, and apoptosis.
• Sample Compatibility:
  • Suitable for serum, plasma, urine, culture media, and other biological fluids.

Sample Preparation

1. Serum Preparation

1. Collect whole blood and allow it to clot at room temperature for 10–20 minutes.
2. Centrifuge at 2,000–3,000 rpm for 20 minutes to separate the serum.
3. Re-centrifuge if precipitates appear during storage.

2. Plasma Preparation

1. Collect whole blood into tubes containing anticoagulants (e.g., EDTA or citrate).
2. Incubate at room temperature for 10–20 minutes.
3. Centrifuge at 2,000–3,000 rpm for 20 minutes to obtain plasma.
4. Re-centrifuge if necessary.

3. Urine Samples

1. Collect urine in aseptic tubes.
2. Centrifuge at 2,000–3,000 rpm for 20 minutes and carefully collect the supernatant.
3. Re-centrifuge if precipitates form during storage.

4. Cell Samples

1. For cell secretions, collect culture supernatants in aseptic tubes and centrifuge at 2,000–3,000 rpm for 20 minutes.
2. For intracellular components:
   • Dilute cells in PBS (pH 7.2–7.4) at 1×10⁷/mL.
   • Disrupt cells via repeated freeze-thaw cycles.
   • Centrifuge at 2,000–3,000 rpm for 20 minutes and collect the supernatant.

5. Tissue Samples

1. Cut and weigh tissue samples, then freeze in liquid nitrogen and store at -80°C.
2. Homogenize samples in PBS (pH 7.4) at 4°C.
3. Centrifuge at 2,000–3,000 rpm for 20 minutes and collect the supernatant.

Important Notes

1. Perform ELISA assays as soon as possible after sample collection.
2. Store samples at -20°C if immediate testing is not feasible.
3. Avoid repeated freeze-thaw cycles to preserve sample integrity.
4. Do not use samples containing sodium azide (NaN₃) as it inhibits HRP activity.

Applications

• Inflammation Research:
  • Study ICE’s role in the activation of pro-inflammatory cytokines such as IL-1beta.
• Apoptosis Studies:
  • Investigate ICE (Caspase-1) activity in programmed cell death and pyroptosis.
• Cancer Research:
  • Explore ICE’s involvement in tumor progression and immune evasion.
• Infectious Disease Studies:
  • Assess ICE’s role in immune responses to bacterial and viral infections.
• Therapeutic Development:
  • Evaluate ICE as a potential target for anti-inflammatory and immune-modulating therapies.

Advantages

1. High Sensitivity and Specificity:
   • Detects low levels of ICE with minimal cross-reactivity or interference.
2. Wide Sample Compatibility:
   • Validated for serum, plasma, urine, cell culture supernatants, and tissue homogenates.
3. Ease of Use:
   • Pre-coated plates and ready-to-use reagents simplify assay execution.
4. Reproducibility:
   • Ensures consistent and reliable results across multiple experiments.

Storage and Stability

• Store all kit components at 2–8°C to maintain reagent integrity.
• Avoid repeated freeze-thaw cycles to ensure assay reliability.

The Human Interleukin 1beta Converting Enzyme (ICE) ELISA Kit is a critical tool for researchers studying inflammation, apoptosis, and related pathways. Its precision, ease of use, and broad compatibility make it indispensable for clinical and experimental applications.