This assay recognizes recombinant and natural human ANX-I.

Human Annexin I (ANX-I) ELISA Kit


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Background

Annexin I belongs to a family of Ca2+-dependent phospholipid-binding proteins that have a molecular weight of approximately 35,000 to 40,000 and are preferentially located on the cytosolic face of the plasma membrane. Annexin I protein has an apparent relative molecular mass of 40 kDa, with phospholipase A2 inhibitory activity. Annexin I is an intracellular protein in search of a function. The low concentrations found in extracellular fluids are the result of a selective secretion. In some tissues annexin I exceeds 0.4% of the total protein. Annexin I is a good substrate of receptor- or non-receptor tyrosine kinases, interacts calcium-dependently with protein kinase C (PKC), and is phosphorylated by this kinase.

Features

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to ANX-I. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for ANX-I and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain ANX-I, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ANX-I in the samples is then determined by comparing the O.D. of the samples to the standard curve.

< SPECIFICITY>
This assay recognizes recombinant and natural human ANX-?. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
15.6 ng/ml-1000 ng/ml.
The standard curve concentrations used for the ELISA’s were 1000ng/ml, 500 ng/ml, 250 ng/ml, 125 ng/ml, 62.5 ng/ml, 31.2 ng/ml, 15.6 ng/ml.

<SENSITIVITY>
The minimum detectable dose of human ANX-? is typically less than 3.9 ng/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Composition

   Reagent Quantity
1. Assay plate 1
2. Standard 2
3. Sample Diluent 1 x 20 ml
4. Biotin-antibody Diluent 1 x 10 ml
5. HRP-avidin Diluent 1 x 10 ml
6. Biotin-antibodyt 1 x 120μl
7. HRP-avidin 1 x 120μl
8. Wash Buffer 1 x 20 ml (25×concentrate)
9. TMB Substrate 1 x 10 ml
10. Stop Solution 1 x 10 ml
Product List

Product Name Cat# Quantity Price

Species : 15.6 ng/ml - 1000 ng/ml.
Assay range : 100 µL (Culture Supernatant, Plasma, Serum)
Assay sample volume : 6-8 hr.+3 hr

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