Angiopoietin-2 (ANGPT2) is a protein that plays a crucial role in angiogenesis, the formation of new blood vessels. It is a ligand for the Tie2 receptor and functions as a regulator of vascular remodeling and endothelial cell function. ANGPT2 is involved in various physiological and pathological processes, including inflammation, cancer, and cardiovascular diseases. ELISA kits for ANGPT2 are used to measure its levels in biological samples such as serum, plasma, and cell culture supernatants.

Overview of ANGPT2 ELISA Kits

• Purpose: To detect and quantify ANGPT2 levels in biological samples. This measurement is useful for research into vascular biology, tumor growth, inflammatory conditions, and cardiovascular diseases.
• Assay Principle: Typically a sandwich ELISA. The assay involves capturing ANGPT2 with a specific antibody immobilized on a microtiter plate, followed by detection using a secondary antibody conjugated to an enzyme (usually HRP). The enzymatic reaction produces a color change that is proportional to the amount of ANGPT2 present.
• Sensitivity: High sensitivity, often in the picogram per milliliter (pg/mL) range, to detect low levels of ANGPT2.
• Specificity: High specificity for ANGPT2, minimizing cross-reactivity with other angiopoietins or proteins.
• Sample Volume: Typically requires 50-100 µL of sample per well.

Content of a Standard ANGPT2 ELISA Kit

• Microtiter Plate: Pre-coated with an anti-ANGPT2 antibody.
• Standards: Recombinant ANGPT2 protein at various concentrations to generate a standard curve.
• Detection Antibody: Biotinylated or enzyme-conjugated anti-ANGPT2 antibody.
• Enzyme Conjugate: Typically streptavidin-HRP, used to amplify the detection signal.
• Substrate Solution: Usually TMB (3,3',5,5'-Tetramethylbenzidine), which produces a color change upon reaction with HRP.
• Stop Solution: Often sulfuric acid or hydrochloric acid, which halts the enzymatic reaction and stabilizes the color change.

Key Steps in the Assay Procedure

• Sample/Standard Addition: Standards and samples are added to wells pre-coated with an anti-ANGPT2 antibody.
• Incubation: The plate is incubated to allow ANGPT2 to bind to the capture antibody.
• Washing: Wells are washed to remove unbound components.
• Detection Antibody Addition: A detection antibody specific to ANGPT2 is added, binding to the captured protein.
• Washing: Excess detection antibody is removed by additional washing.
• Enzyme Conjugate Addition: Enzyme-conjugated secondary antibody binds to the detection antibody.

Applications

• Research: Investigating the role of ANGPT2 in angiogenesis, vascular remodeling, and various diseases such as cancer, cardiovascular disorders, and inflammatory conditions.
• Clinical Diagnostics: Measuring ANGPT2 levels to understand its involvement in disease processes, assess disease progression, and evaluate therapeutic responses.

ANGPT2 ELISA kits are valuable tools for understanding vascular biology and disease mechanisms. They enable researchers and clinicians to measure ANGPT2 levels in biological samples, providing insights into its role in various physiological and pathological conditions.