Invitrogen™ Rat Insulin ELISA Kit

Invitrogen™ Insulin Rat ELISA Kit - Detailed Description

The Invitrogen™ Insulin Rat ELISA Kit is a highly sensitive sandwich ELISA designed for the quantitative measurement of insulin in rat biological samples, including serum, plasma, and cell culture supernatants. This kit offers a robust and specific method for assessing insulin levels, enabling researchers to study insulin’s role in metabolism, diabetes, and endocrine signaling.

Key Features

1. High Sensitivity and Precision:
   • Analytical Sensitivity: 5 µIU/mL.
   • Assay Range: 4.69–300 µIU/mL.
   • Intraassay CV: <10%.
   • Interassay CV: <12%.
2. Versatile Sample Compatibility:
   • Suitable for rat serum, plasma, and cell culture supernatants.
   • Requires as little as 50 µL per sample for serum and plasma, and 100 µL for supernatants.
3. Comprehensive Kit Design:
   • Includes pre-coated 96-well plates, standards, controls, and all necessary reagents for streamlined workflows.
4. Reliable Sandwich ELISA Format:
   • Uses matched antibody pairs to ensure specific detection of natural and recombinant rat insulin.
5. Time-Efficient Workflow:
   • Hands-on time: 1 hr 20 min.
   • Total time-to-result: 4 hr 45 min.

Applications and Use Cases

1. Diabetes and Metabolic Research:

• Hyperglycemia Studies:
  • Quantify insulin levels in diabetic models to study hyperglycemia and insulin resistance.
• Insulin Secretion Analysis:
  • Measure insulin release from pancreatic beta cells.

2. Endocrinology Research:

• Hormonal Regulation Studies:
  • Investigate insulin’s role in carbohydrate, protein, and lipid metabolism.
• Insulin and Tumor Biology:
  • Study insulin presence in insulinoma and beta-cell-origin tumors.

3. Pharmacological Studies:

• Drug Efficacy Testing:
  • Evaluate the impact of anti-diabetic drugs on insulin levels.
• Therapeutic Development:
  • Analyze insulin-modulating compounds in preclinical studies.

4. Cell and Molecular Biology:

• Cell Culture Studies:
  • Measure insulin secretion in cultured pancreatic islet cells.
• Signal Transduction Pathways:
  • Investigate insulin’s role in metabolic and anabolic signaling.

Kit Components

Assay Principle

This ELISA kit employs a sandwich ELISA method for accurate quantification of rat insulin:

1. Capture Phase:
   • The microplate is pre-coated with a specific antibody to insulin, capturing insulin in the sample.
2. Detection Phase:
   • A biotinylated detection antibody binds to a different epitope of the insulin molecule, forming a sandwich complex.
3. Signal Amplification:
   • Streptavidin-HRP binds to the biotinylated antibody, amplifying the signal.
4. Signal Development:
   • Addition of TMB substrate produces a measurable color change proportional to insulin concentration.
5. Quantification:
   • Measure optical density (OD) at 450 nm, and calculate insulin concentrations using the standard curve.

Performance Specifications

Storage and Shipping

• Storage Conditions: Store all components at 2–8°C.
• Shipping Conditions: Shipped on wet ice to maintain reagent stability.

Target Information: Insulin

• Function:
  • A peptide hormone secreted by pancreatic beta cells, insulin regulates carbohydrate, protein, and lipid metabolism.
  • Promotes glucose uptake, glycogen storage, and protein synthesis while inhibiting gluconeogenesis and lipolysis.
• Structure:
  • Composed of 51 amino acids, processed from proinsulin into two chains (A and B) linked by disulfide bonds.
• Clinical Relevance:
  • Insulin deficiency leads to diabetes mellitus, a leading cause of morbidity.
  • Found in insulinomas, tumors originating from pancreatic beta cells.
• Gene Information:
  • Gene Symbol: Ins1, Ins2.
  • Aliases: Insulin-1, Insulin-2.
  • UniProt ID: P01322, P01323.

Advantages of the Kit

1. High Sensitivity and Precision:
   • Detects very low levels of insulin, ensuring accurate and reliable data.
2. Validated Performance:
   • Rigorously tested for sensitivity, specificity, and reproducibility.
3. User-Friendly Design:
   • Ready-to-use reagents and pre-coated plates simplify assay setup.
4. Broad Applications:
   • Supports research in diabetes, metabolism, and pharmacology.

The Invitrogen™ Insulin Rat ELISA Kit is a reliable, high-sensitivity assay for quantifying insulin in rat biological samples. Its precision, compatibility with multiple sample types, and validated performance make it a critical tool for researchers studying metabolic regulation, diabetes, and endocrine signaling pathways.