β-Defensin 2 ELISA Kit

Name: β-Defensin 2 ELISA Kit
SKU: DEIA040J
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Stool

Detection Method

sELISA

Intended Use

This assay is intended for the quantitative determination of β-defensin 2 in stool. For in vitro diagnostic use only.

Contents of Kit

1.Microtiter plate, pre-coated, 12 x 8wells
2.Wash buffer concentrate 10x, 2 x 100ml
3.Conjugate concentrate, (goat anti-β-defensin 2,HRP-conjugated), 1 x 200μl
4.Standards, lyophilised, 2 x 5 vials
5.Standard dilution buffer, ready-to-use, 1 x 20ml
6.Control 1, lyophilised, (see specification for range), 2 x 1 vial
7.Control 2, lyophilised, (see specification for range), 2 x 1 vial
8.Extraction buffer concentrate 2.5x, 2 x 100ml
9.Substrate (tetramethylbenzidine), ready-to-use, 1 x 15ml
10.Stop solution, ready-to-use, 1 x 15ml

Performance Characteristics

Accuracy – Precision
Repeatability (Intra-Assay); n = 30
The repeatability was assessed with 2 stool samples under constant parameters (same operator, instrument, day and kit lot).

Reproducibility (Inter-Assay); n = 24
The reproducibility was assessed with 4 stool samples under varying parameters(different operators, instruments, days and kit lots).

Accuracy – Trueness
The trueness states the closeness of the agreement between the result of a measurement and the true value of the measurand. Therefore, β-defensin 2 spikes with
known concentrations were added to 8 different stool-samples. In the table below, 4 exemplary samples are shown:

Linearity
The linearity states the ability of a method to provide results proportional to the concentration of analyte in the test sample within a given range. This was assessed according to CLSI guideline EP06-A with a serial dilution of 3 different stool samples.
For β-defensin 2 in stool, the method has been demonstrated to be linear from 0.065 to 2.200ng/ml, showing a non-linear behaviour of less than ± 20% in this interval.

Analytical sensitivity
The following values have been estimated based on the concentrations of the standard without considering possibly used sample dilution factors.
Limit of blank, LoB 0.0085ng/ml
Limit of detection, LoD 0.0155ng/ml
Limit of quantitation, LoQ 0.0232ng/ml
The evaluation was performed according to the CLSI guideline EP-17-A2. The specified accuracy goal for the LoQ was 20 % CV.
Analytical specificity
The specificity of the antibody was tested by measuring the cross-reactivity against a range of compounds with structural similarity to β-defensin 2. There was no crossreactivity observed.

General Description

The β-defensins are an integral part of the congenital immune system and contribute with their antimicrobial effect to the barrier function of intestinal epithelial cells.
Defensins exert a variable degree of antimicrobial activity against bacteria, fungi, and some enveloped viruses. Vertebrate defensins are classified as α- or β-defensins, based on their pattern of disulfide bridges. Nine human defensins of epithelial origin have been found, three of them being β-defensins (HBD-1, -2 and -3). The expression of β-defensins is induced by the pro-inflammatory cytokines and also through microorganisms (e.g. E. coli, H. pylori or P. aeruginosa).
A β-defensin 2 deficiency can, for example, be observed in the intestinal mucous of patients with Crohn's disease. The defense system of the mucous membrane is therefore restricted and allows an increased invasion of bacteria, which could possibly lead to a typical infection in Crohn's disease patients. Whether the β-defensin 2 deficiency could even play a role in the development of Crohn's disease is currently being researched. As is the possibility that it is the probiotic bacterium, which produces β-defensin.
Indications
• Reduced β-defensin levels with Crohn's disease (HBD-2)
• Increased β-defensin levels with Colitis Ulcerosa (HBD-2)

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Transcriptional landscape of psoriasis identifies the involvement of IL36 and IL36RN

BMC GENOMICS

Authors: Keermann, Maris; Koeks, Sulev; Reimann, Ene; Prans, Ele; Abram, Kristi; Kingo, Kuelli

Abstract

Background: In present study we performed whole transcriptome analysis in plaque psoriasis patients and compared lesional skin with non-lesional skin and with the skin from healthy controls. We sequenced total RNA from 12 lesional (LP), 12 non-lesional (NLP) and from 12 normal (C) skin biopsies. Results: Compared with previous gene expression profiling studies we had three groups under analysis - LP, NLP and C. Using NLP samples allows to see the transcriptome of visually normal skin from psoriasis patient. In LP skin S100A12, S100A7A, LCE3E, DEFB4A, IL19 were found up regulated. In addition to already these well-described genes, we also found several other genes related to psoriasis. Namely, KLK9, OAS2, OAS3, PLA2G, IL36G, IL36RN were found to be significantly and consistently related to the psoriatic lesions and this finding is supported also by previous studies. The genes up-regulated in the LP samples were related to the innate immunity, IL17 and IL10 networks. In NLP samples innate immunity and IL17 network were activated, but activation of IL10 network was not evident. The transcriptional changes characteristic in the NLP samples can be considered as a molecular signature of "dormant psoriasis". Conclusions: Taken together, our study described the transcriptome profile characteristic for LP and NLP psoriatic skin. RNA profile of the NLP skin is in between the lesional and healthy skin, with its own specific pattern. We found that both LP and NLP have up-regulated IL17 network, whereas LP skin has up regulated IL10 related cytokines (IL19, IL20, IL24). Moreover, IL36G and IL36RN were identified as strong regulators of skin pathology in both LP and NLP skin samples, with stronger influence in LP samples.

Tissue-engineered human psoriatic skin supplemented with cytokines as an in vitro model to study plaque psoriasis

REGENERATIVE MEDICINE

Authors: Pouliot-Berube, Claudia; Zaniolo, Karine; Guerin, Sylvain L.; Pouliot, Roxane

Abstract

Aim: Psoriasis is a chronic inflammatory skin disease. To study its complex etiology, a psoriatic skin substitute model supplemented with a cytokine cocktail has been used. Materials & methods: Reconstructed psoriatic skin substitutes were supplemented with a cocktail of four cytokines: TNF-alpha, IL-1 alpha, IL-6 and IL-17A, to monitor their impact on gene expression by DNA microarray. Results: Gene profiling analyses identified several deregulated genes reported as being also deregulated in psoriasis skin in vivo (S100A12, IL-8, DEFB4A and KYNU). The expression of those genes was dramatically increased compared with basal levels of controls (p < 0.005 to < 0.05). Conclusion: Psoriatic substitutes supplemented with a cocktail of TNF-alpha, IL-1 alpha, IL-6 and IL-17A showed similar transcriptome alterations to those found in psoriasis.