Mouse Angiotensin II EasyTest™ EIA Kit

Product Features

Product Name            Mouse Angiotensin II EasyTest™ EIA Kit
Assay Type                   Solid Phase Competitive ELISA
Format                           96-well strip plate
Assay Process             Two wash step, three incubations
Assay Length              2.0hours
Sample Volume         Serum , Plasma, 50µL
Detection Method    Colorimetric
Sensitivity                   6pg/mL
Assay Range                0.49-2000pg/mL
Reactivity                     Mouse
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               AGT; AI265500; AngI; AngII; Aogen; Serpina8
Gene ID                         11606
Swissprot ID               P11859

Standard Curve       Mouse Angiotensin II EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-ANGII Antibody
• Standard ANGII peptide
• Biotinylated ANGII peptide
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 25 µl of anti-ANGII antibody to all wells.
3. Add 75 µl of 1 x assay diluent to the blank wells and 50 µl to B0 wells.
4. Add 50 µl of standard peptide/samples to the appropriate wells.
5. Add 25 µl of biotinylated peptide into each well except the blank wells.
6. Incubate 1.0 h at RT.
7. Wash and add 100 µl of diluted streptavidin-HRP solution to all wells.
8. Incubate 45 min at RT.
9. Wash and add 100 µl of TMB Substrate Solution into each well.
10. Incubate 15 min at RT.
11. Add 50 µl of Stop Solution to each well.
12. Read at 450 nm/570 nm.

Application

In blood, Angiotensin II binds with big proteins. Acidification of blood is required to separate and purify Angiotensin II.

Background

Angiotensinogen is an α-2-globulin produced constitutively and released into the circulation mainly by the liver. It is a member of the serpin family. Angiotensin I is converted to angiotensin II (AII) through removal of two C-terminal residues by the enzyme angiotensin-converting enzyme (ACE), primarily through ACE within the lung . ACE found in other tissues of the body has no physiological role. Angiotensin II acts as an endocrine, autocrine/paracrine, and intracrine hormone. Angiotensin II is degraded to angiotensin III by angiotensinases located in red blood cells and the vascular beds of most tissues.

Angiotensin II increases blood pressure by stimulating the Gq protein in vascular smooth muscle cells. In addition, angiotensin II acts at the Na/H exchanger in the proximal tubules of the kidney to stimulate Na reabsorption and H excretion which is coupled to bicarbonate reabsorption. This ultimately results in an increase in blood volume, pressure, and pH.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Mouse Angiotensin II EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS