Mouse ACE ELISA Kit

Catalog No.: BEK1277

Size: 96T

Range: 93.8 pg/ml - 6000 pg/ml

Sensitivity < 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of ACE in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Angiotensin-converting enzyme (ACE) is a circulating enzyme that participates in the body's renin-angiotensin system (RAS), which mediates extracellular volume and arterial vasoconstriction. It is secreted by pulmonary and renal endothelial cells and catalyzes the conversion of decapeptide angiotensin I to octapeptide angiotensin II. It plays an important role in blood pressure regulation, and also plays a crucial role in fertilization through its GPIase activity. The enzyme is also able to inactivate bradykinin, a potent vasodilator. It has a molecular weight of 80,073.

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- ACE polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- ACE polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the ACE amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ACE can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse ACE antibody
2. Lyophilized mouse ACE standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-mouse ACE antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml
11. Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L