Human High-temperature requirement factor A3 ELISA kit

Name: Human High-temperature requirement factor A3 ELISA kit
SKU: DEIA-BJ1053
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

Serum, tissue, cell culture supernatants

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The ELISA Kit from CD provides a highly sensitive and specific quantitative determination of HtrA3 in serum, tissue (Placenta) and cell culture supernatants. For research use only, not for use in diagnostic procedures.

Contents of Kit

1. Microtiter plate: The plate contains 6×16 strips coated with monoclonal anti-HtrA3.
2. Assay Buffer 2.5: Bottle contains 25 ml buffer with additives.
3. Wash Buffer Concentrate: Two bottles containing each 25 ml of a buffer concentrate. It has to be diluted 20-fold with distilled or deionised water before use.
4. HtrA3 Standard (lyophilised): The vial contains 200 ng of lyophilized HtrA3. It has to be reconstituted with 1 ml of HtrA3 Standard Buffer prior to use to get a 200 ng/ml standard solution.
5. HtrA3 Standard Buffer: Solution to reconstitute HtrA3 lyophilized.
6. Serum Standard Diluent: This solution (20 ml) contains certified human serum in a buffer with additives. Ready to use.
Warning: Component contains human serum! Wear eye, hand, face and clothing protection when using this material!
7. Detection Buffer: Solution (25 ml) consists of phosphate buffer containing additives.
8. Biotinylated Antibody: Solution (150 μl) contains biotinylated monoclonal anti-HtrA3 antibody in buffer with additives. The antibody has to be diluted 100-fold with Detection Buffer before use.
9. Conjugate Solution: This solution (300 μl) contains a highly polymeric streptavidin-peroxidase conjugate with preservatives. It has to be diluted 100-fold with Detection Buffer prior to use.
10. TMB Substrate: Bottle contains 12 ml of a TMB solution ready to use.
11. Stop Solution: Bottle contains 12 ml 0.25 M H₂SO₄. Ready to use.
Warning: Stop Solution contains 0.25 M sulfuric acid. Wear eye, hand, face and clothing protection when using this material!

Storage

1. All components of the kit can be stored in the refrigerator (2-8°C).
2. Note! Wash buffer concentrate can precipitate in refrigerator. We recommend stored at 18-35°C
3. HtrA3 Standard Buffer should be used immediately or stored at -20°C.
4. Once reconstituted, the HtrA3 Standard should be used immediately or stored at -80 °C.
5. Diluted Biotinylated antibody and diluted Conjugate solution should be prepared freshly directly before use. When running a partial plate, only suitable aliquots of these solutions should be made.

Precision

1. Within assay precision
The within assay precision was measured by assaying four control samples in 10 duplicates in Assay Buffer on one plate.

2. Between assay precision
The between assay variation was measured by assaying.

Sensitivity

The sensitivity was determined up to 700 pg/ml.

General Description

Human HtrA3 (high temperature requirement A3) is a serine protease of the HtrA family. HtrA3 was initially identified in the developing placenta both in mouse and in human as a serine protease associated with pregnancy. HtrA3 is now known to inhibit trophoblast invasion during placental development, and regulate ovarian development, granulosa cell differentiation and luteinisation. Studies in mice have also suggested that HtrA3 inhibits TGF-β signalling during embryo development. It has two isoforms [long (HtrA3-L) and short (HtrA3-S)] and recently, HtrA3 was identified as a potential diagnostic marker for early detection of preeclampsia, a life-threatening pregnancy-specific disorder.

Standard Curve

Typical assay data and Typical standard curve (prepared with Assay Buffer 2.5).

Typical assay data and Typical standard curve (prepared with Serum Standard Diluent).

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Immune response against HtrA proteases in children with cutaneous mastocytosis

ACTA BIOCHIMICA POLONICA

Authors: Renke, Joanna; Kedzierska-Mieszkowska, Sabina; Lange, Magdalena; Nedoszytko, Boguslaw; Liberek, Anna; Plata-Nazar, Katarzyna; Renke, Marcin; Wenta, Tomasz; Zurawa-Janicka, Dorota; Skorko-Glonek, Joanna; Lipinska, Barbara

Abstract

Mast cells play an important role in both, the innate and adaptive immunity, however, clonal proliferation of abnormal mast cells in various organs leads to mastocytosis. A skin variant of the disease, cutaneous mastocytosis (CM) is the most frequent form of mastocytosis in children. HtrA proteases are modulators of important cellular processes, including cell signaling and apoptosis, and are related to development of several pathologies. The above and the observation that mast cells constitutively release the HtrA1 protein, prompted us to investigate a possible involvement of the HtrA proteins in pediatric CM. Levels of the serum autoantibodies (IgG) against the recombinant HtrA proteins (HtrA1-4) in children with CM (n=36) and in healthy controls (n=62) were assayed. Anti-HtrA IgGs were detected using enzyme linked immunosorbent assay (ELISA) and Western-blotting. In the CM sera, levels of the anti-HtrA1 and anti-HtrA3 autoantibodies were significantly increased when compared to the control group, while the HtrA protein levels were comparable. No significant differences in the anti-HtrA2 IgG level were found; for the anti-HtrA4 IgGs lower levels in CM group were revealed. In healthy children, the IgG levels against the HtrA1, -3 and -4 increased significantly with the age of children; no significant changes were observed for the anti-HtrA2 IgG. Our results suggest involvement of the HtrA1 and HtrA3 proteins in pediatric CM; involvement of the HtrA4 protein is possible but needs to be investigated further. In healthy children, the autoantibody levels against HtrA1, -3 and -4, but not against HtrA2, increase with age.

DNA methylation-regulated microRNA pathways in ovarian serous cystadenocarcinoma: A meta-analysis

COMPUTATIONAL BIOLOGY AND CHEMISTRY

Authors: Agustriawan, David; Huang, Chien-Hung; Sheu, Jim Jinn-Chyuan; Lee, Shan-Chih; Tsai, Jeffrey J. P.; Kurubanjerdjit, Nilubon; Ng, Ka-Lok

Abstract

Epigenetic regulation has been linked to the initiation and progression of cancer. Aberrant expression of microRNAs (miRNAs) is one such mechanism that can activate or silence oncogenes (OCGs) and tumor suppressor genes (TSGs) in cells. A growing number of studies suggest that miRNA expression can be regulated by methylation modification, thus triggering cancer development. However, there is no comprehensive in silico study concerning miRNA regulation by direct DNA methylation in cancer. Ovarian serous cystadenocarcinoma (OSC) was therefore chosen as a tumor model for the present work. Twelve batches of OSC data, with at least 35 patient samples in each batch, were obtained from The Cancer Genome Atlas (TCGA) database. The Spearman rank correlation coefficient (SRCC) was used to quantify the correlation between the CpG DNA methylation level and miRNA expression level. Meta analysis was performed to reduce the effects of biological heterogeneity among different batches. MiRNA-target interactions were also inferred by computing SRCC and meta-analysis to assess the correlation between miRNA expression and cancer-associated gene expression and the interactions were further validated by a query against the miRTarBase database. A total of 26 potential epigenetic-regulated miRNA genes that can target OCGs or TSGs in OSC were found to show biological relevance between DNA methylation and miRNA gene expression. Furthermore, some of the identified DNA-methylated miRNA genes; for instance, the miR-200 family, were previously identified as epigenetic-regulated miRNAs and correlated with poor survival of ovarian cancer. We also found that several miRNA target genes, BTG3, NDN, HTRA3, CDC25A, and HMGA2 were also related to the poor outcomes in ovarian cancer. The present study proposed a systematic strategy to construct highly confident epigenetic-regulated miRNA pathways for OSC. The findings are validated and are in line with the literature. The inclusion of direct DNA methylated miRNA events may offer another layer of explanation that along with genetics can give a better understanding of the carcinogenesis process. (C) 2016 Elsevier Ltd. All rights reserved.