Monkey Apolipoprotein A1, ApoA1 ELISA Kit

Catalog No.: BZEK2873

Size: 96T

Range: 0.25 μg/ml - 4 μg/ml

Sensitivity: 0.1 μg/ml

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of ApoA1 in monkey serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Apolipoprotein AI (ApoA-I) is a protein that in humans is encoded by the APOA1 gene. As the major component of HDL particles, it has a specific role in lipid metabolism. APOA1 is located on chromosome 11, with its specific location being 11q23-q24. The gene contains 4 exons. The encoded apolipoprotein A-I, is a 28.1 kDa protein composed of 243 amino acids; 21 peptides have been observed through mass spectrometry data. Apolipoprotein AI is the major protein component of high density lipoprotein (HDL) particles in plasma. Chylomicrons secreted from the intestinal enterocyte also contain apo-AI, but it is quickly transferred to HDL in the bloodstream. The protein, as a component of HDL particles, enables efflux of fat molecules by accepting fats from within cells (including macrophages within the walls of arteries which have become overloaded with ingested fats from oxidized LDL particles) for transport (in the water outside cells) elsewhere, including back to LDL particles or to the liver for excretion.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- ApoA1 antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- ApoA1 antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 ApoA1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ApoA1 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti- monkey ApoA1 antibody

2. Standard: 0.5ml (8 μg/ml)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (30×): 20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti- monkey ApoA1 antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L