Intended use
This immunoassay kit allows for the in vitro quantitative determination of General LTAD concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.
Test principle
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General LTAD, During the reaction, General LTAD in the sample or standard competes with a fixed amount of biotin-labeled General LTAD for sites on a pre-coated Monoclonal antibody specific to General LTAD. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. 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 General LTAD in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Lipoteichoic acid and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	99-111%	101-113%	91-103%	87-95%
EDTA plasma(n=5)	103-115%	83-93%	94-103%	110-120%
heparin plasma(n=5)	105-114%	100-109%	92-101%	100-110%

 

General Annotation

 

Function:

LTA may bind to target cells non-specifically through membrane phospholipids, or specifically to CD14 and to Toll-like receptors. Binding to TLR-2 has shown to induce NF-κB expression(a central transcription factor), elevating expression of both pro- and anti-apoptotic genes. Its activation also induces mitogen-activated protein kinases (MAPK) activation along with Phosphoinositide 3-kinase activation.

 

Location:

Lipoteichoic acid (LTA) is a major constituent of the cell wall of Gram-positive bacteria. These organisms have an inner (or cytoplasmic) membrane and, external to it, a thick (up to 80 nanometer) peptidoglycan layer. The structure of LTA varies between the different species of Gram positive bacteria and may contain long chains of ribitol or glycerol phosphate. LTA is anchored to the cell membrane via a glyceride. It acts as regulator of autolytic wall enzymes (muramidases). It has antigenic properties being able to stimulate specific immune response and is released from the bacterial cells mainly after bacteriolysis induced by lysozyme, cationic peptides from leucocytes, or beta-lactam antibiotics.

 

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