Mouse Epiregulin ELISA kit

Name: Mouse Epiregulin ELISA kit
SKU: DEIA5341
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

Serum, plasma, and cell culture supernatants

Species Reactivity

Mouse

Detection Method

sELISA

Intended Use

The Mouse Epiregulin ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse Epiregulin in serum (mouse Epiregulin concentration is low in normal serum/plasma and may not be detectable in this assay), plasma and cell culture supernatants.

Contents of Kit

1. Epiregulin Microplate (Item A): 96 wells (12 strips × 8 wells) coated with anti-Mouse Epiregulin. Store at 4°C for 1 month. (Return unused wells to the pouch containing desiccant pack, reseal along entire edge.)
2. Wash Buffer Concentrate (20×) (Item B): 25 ml of 20× concentrated solution. Store at 4°C for 1 month.
3. Standard Protein (Item C): 2 vials of Mouse Epiregulin. 1 vial is enough to run each standard in duplicate. Store at -80°C for 1 week.
4. Detection Antibody Epiregulin (Item F): 2 vials of biotinylated anti-Mouse Epiregulin. Each vial is enough to assay half the microplate. Store at 4°C for 5 days.
5. HRP-Streptavidin Concentrate (Item G): 200 μl 300× concentrated HRP-conjugated streptavidin. Do not store and reuse.
6. TMB One-Step Substrate Reagent (Item H): 12 ml of 3,3,5,5'-tetramethylbenzidine (TMB) in buffer solution.
7. Stop Solution (Item I): 8 ml of 0.2 M sulfuric acid.
8. Assay Diluent A (Item D): 30 ml of diluent buffer, 0.09% sodium azide as preservative.
9. Assay Diluent B (Item E): 15 ml of 5× concentrated buffer. Store at 4°C for 1 month.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C. For prepared reagent storage, see "Reagents And Materials Provided".

Precision

Intra-Assay CV%: <10%
Inter-Assay CV%: <12%

Detection Range

10 pg/ml - 6000 pg/ml

Sensitivity

The minimum detectable dose of Mouse Epiregulin was determined to be 10 pg/ml. Minimum detectable dose is defined as the analyte concentration resulting in an absorbance that is 2 standard deviations higher than that of the blank (diluent buffer).

Standard Curve

These standard curves are for demonstration only. A standard curve must be run with each assay.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Biomarkers of benefit from cetuximab-based therapy in metastatic colorectal cancer: interaction of EGFR ligand expression with RAS/RAF, PIK3CA genotypes

BMC CANCER

Authors: Pentheroudakis, George; Kotoula, Vassiliki; De Roock, Wendy; Kouvatseas, George; Papakostas, Pavlos; Makatsoris, Thomas; Papamichael, Demetris; Xanthakis, Ioannis; Sgouros, Joseph; Televantou, Despina; Kafiri, Georgia; Tsamandas, Athanassios C.; Razis, Evangelia; Galani, Eleni; Bafaloukos, Dimitrios; Efstratiou, Ioannis; Bompolaki, Iliada; Pectasides, Dimitrios; Pavlidis, Nicholas; Tejpar, Sabine; Fountzilas, George

Abstract

Background: More than half of patients with KRAS-wild type advanced colorectal cancer (CRC) fail anti-EGFR monoclonal antibodies. We studied EGFR-axis messenger RNA (mRNA) expression and RAS, RAF, PIK3CA mutations in order to identify additional biomarkers of cetuximab efficacy. Methods: Previously genotyped (KRAS, NRAS, BRAF, PIK3CA mutations) formalin-fixed paraffin-embedded tumour biopsies of 226 cetuximab-treated CRC patients (1st to 3rd line therapy) were assessed for mRNA expression of epidermal growth factor receptor (EGFR) and its ligands EGF, Transofrming Growth Factor-a (TGFA), Amphiregulin (AREG) and Epiregulin (EREG) with real time quantitative PCR. Mutations were detected in 72 (31.9%) tumours for KRAS, in 6 (2.65%) for BRAF, in 7 (3.1%) for NRAS and in 37 (16.4%) for PIK3CA. Results: Only PIK3CA mutations occasionally coexisted with other gene mutations. In univariate analysis, prognostic significance for survival (from metastases until death) was seen for BRAF mutations (Hazard Ratio HR 8.1, 95% CI 3.4-19), codon 12-only KRAS mutations (HR 1.62, 95% CI 1.1-2.4), high AREG mRNA expression only in KRAS wild type CRC (HR 0.47, 95% CI 0.3-0.7) and high EREG mRNA expression irrespective of KRAS mutation status (HR 0.45, 95% CI 0.28-0.7). EREG tumoural mRNA expression was significantly associated with a 2.26-fold increased likelihood of objective response to cetuximab therapy (RECIST 1.1). In multivariate analysis, favourable predictive factors were high AREG mRNA in KRAS wild type tumours, high EREG mRNA, low Ephrin A2 receptor mRNA. Cetuximab-treated patients with AREG-low KRAS wild type CRC fared very poorly, their survival being similar to KRAS mutant CRC. Patients with KRAS codon 13 or other non-codon 12 mutations had a median survival (30 months, 95% CI 20-35) similar to that of patients with KRAS wild-type (median survival 29 months, 95% CI 25-35), in contrast to patients with KRAS codon 12 mutations who fared worse (median survival 19 months, 95% CI 15-26). Conclusions: BRAF and codon 12 KRAS mutations predict for adverse outcome of CRC patients receiving cetuximab. AREG mRNA reflects EGFR signalling in KRAS wild type tumours, predicting for cetuximab efficacy when high and failure when low. EREG may have a prognostic role independent of KRAS mutation.

Does Bone Morphogenetic Protein 6 (BMP6) Affect Female Fertility in the Mouse?

BIOLOGY OF REPRODUCTION

Authors: Sugiura, Koji; Su, You-Qiang; Eppig, John J.

Abstract

Bone morphogenetic protein 6 (BMP6) is a transforming growth factor beta superfamily member produced by mammalian oocytes as well as other cell types. Despite well-characterized effects of recombinant BMP6 on granulosa cells in vitro, the function of BMP6 in vivo has been ill-defined. Therefore, the effects of genetic deletion of the Bmp6 gene on female mouse fertility were assessed. The mean litter size of Bmp6(-/-) females was reduced by 22% (P < 0.05) compared to Bmp6(+/+) controls. Not only did Bmp6-/- females naturally ovulate 24% fewer eggs, but competence of in vitro-matured oocytes to complete preimplantation development after fertilization in vitro was decreased by 50%. No apparent effect of Bmp6 deletion on either the morphology or the dynamics of follicular development was apparent. Nevertheless, levels of luteinizing hormone (LH)/human chorionic gonadotropin (hCG)-induced transcripts, which encode proteins required for cumulus expansion (HAS2, PTGS2, PTX3, and TNFAIP6), and of epidermal growth factor-like peptides (AREG, BTC, and EREG) were lower in Bmp6(-/-) mice than in controls after administration of a reduced dose of hCG (1 IU) in vivo. LH receptor (Lhcgr) transcript levels were not significantly lower in Bmp6(-/-) granulosa cells, suggesting that BMP6 is required for processes downstream of LH receptors. To assess whether another oocyte-derived BMP, BMP15, could have BMP6-redundant functions in vivo, the fertility of Bmp15/Bmp6 double mutants was assessed. Fertility was not significantly reduced in double-homozygous mutants compared with that in double-heterozygous controls. Therefore, BMP6 promotes normal fertility in female mice, at least in part, by enabling appropriate responses to LH and normal oocyte quality. Thus, Bmp6 probably is part of the complex genetic network that determines female fertility.