Invitrogen™ Human Haptoglobin ELISA Kit

The Invitrogen™ Human Haptoglobin ELISA Kit provides an advanced and reliable approach to quantitatively measure haptoglobin levels in various biological samples such as plasma, serum, and other fluids. This kit is widely used in research to explore the role of haptoglobin in acute-phase responses, inflammation, hemolysis, and liver-related diseases.

Key Features:

1. High Sensitivity and Range:
   • Analytical Sensitivity: Detects as low as 4.69 ng/mL, enabling identification of even minimal haptoglobin concentrations.
   • Dynamic Range: Measures concentrations between 7.81 ng/mL and 500 ng/mL, supporting diverse research requirements.
2. Rapid Assay Time:
   • Requires only 1 hr 20 min hands-on time.
   • Provides results within 3 hr 30 min for efficient workflow management.
3. Precision:
   • Intraassay Coefficient of Variation (CV): <10%
   • Interassay CV: <10%
4. User-Friendly Design:
   • The kit includes a pre-coated 96-well microplate, eliminating the need for additional preparation and streamlining experimental setup.
5. Flexible Applications:
   • Compatible with plasma (100 µL) and serum (100 µL) samples.
   • Suitable for use with a colorimetric microplate reader, ensuring compatibility with most laboratory setups.

Kit Components:

• Pre-coated 96-Well Microplate: Antibodies specific to haptoglobin are pre-coated for target capture.
• Haptoglobin Standard: For generating a standard curve to determine unknown sample concentrations.
• Biotinylated Detection Antibody: Specific for haptoglobin, labeled with biotin for subsequent detection.
• HRP Conjugate: Enhances detection by binding to the biotin-labeled antibody.
• Buffers and Reagents:
  • Standard & Sample Diluent
  • Biotinylated Detection Antibody Diluent
  • HRP Conjugate Diluent
  • Wash Buffer
  • TMB Substrate for colorimetric detection.
  • Stop Solution to halt the enzyme-substrate reaction.
• Accessories: Plate sealer for secure handling during incubation steps.

Principle of the Assay:

The Human Haptoglobin ELISA Kit uses a sandwich ELISA technique:

1. Capture Phase:
   • The pre-coated microplate contains antibodies that specifically bind to haptoglobin in the sample.
   • Samples or standards are added to the wells, allowing haptoglobin to bind to the capture antibodies.
2. Detection Phase:
   • A biotinylated detection antibody binds to a different epitope of haptoglobin, forming a "sandwich" complex.
   • HRP-conjugated avidin binds to the biotinylated antibody.
3. Signal Development:
   • TMB substrate reacts with the HRP enzyme to produce a colorimetric signal proportional to haptoglobin concentration.
   • Stop solution halts the reaction, and the absorbance is measured using a microplate reader.
4. Quantification:
   • A standard curve generated from the provided standards is used to determine the concentration of haptoglobin in unknown samples.

Applications:

1. Inflammation and Acute Phase Response:
   • Study haptoglobin as an acute-phase reactant during inflammation or infection.
2. Hemolysis and Red Blood Cell Turnover:
   • Measure decreased haptoglobin levels associated with conditions like sickle cell anemia, thalassemia, and autoimmune hemolytic anemia.
3. Liver Disease Research:
   • Assess haptoglobin levels in liver dysfunction, where synthesis may be impaired.
4. Oxidative Stress Studies:
   • Investigate haptoglobin's role in scavenging free hemoglobin and reducing oxidative damage.
5. Immune Response Monitoring:
   • Explore haptoglobin's involvement in immune modulation and its interaction with the complement system.

Haptoglobin Overview:

Haptoglobin is an α2-globulin plasma protein synthesized in the liver. It plays a critical role in:

• Scavenging free hemoglobin during hemolysis to prevent oxidative damage.
• Acting as an acute-phase protein, with levels increasing in response to inflammation, infection, or injury.
• Serving as a biomarker for various conditions such as anemia, inflammation, and liver diseases.

Protein Details:

• Aliases: BP, HP2ALPHA2, HPA1S, Zonulin
• Structure: Comprised of two alpha and two beta chains, forming three major phenotypes: Hp 1-1, Hp 2-1, Hp 2-2.
• Gene Symbol: HP (Human Gene ID: 3240)
• UniProt ID: P00738

Validation and Quality Control:

Each lot of the Human Haptoglobin ELISA Kit undergoes rigorous testing to ensure:

• Sensitivity and specificity for haptoglobin.
• Precision and reproducibility across different lots.
• Consistent performance in laboratory conditions.

Shipping and Storage:

• Shipping: Shipped on wet ice to maintain stability during transit.
• Storage: Refer to specific documentation for storage details; typically stored at 2–8°C.

Advantages of the Kit:

1. Ease of Use: Pre-coated microplates and ready-to-use reagents streamline the assay process.
2. Rapid Results: Obtain results in under 4 hours.
3. Wide Dynamic Range: Suitable for both high and low concentrations of haptoglobin.
4. Versatile Applications: Ideal for various research fields, including immunology, hematology, and liver disease studies.

Important Notes:

• For research use only. Not intended for diagnostic or therapeutic applications.
• This product is not automated and is designed for professional use in controlled laboratory environments.

The Invitrogen™ Human Haptoglobin ELISA Kit is an indispensable tool for researchers studying inflammatory processes, hemolytic disorders, and liver function. With its high sensitivity, precision, and ease of use, this kit supports a wide range of biomedical investigations.