Statistik und Sichtungsnachweis dieser Seite findet sich am Artikelende
| [1.] Iam/Fragment 046 01 - Diskussion Zuletzt bearbeitet: 2014-03-12 20:16:39 Graf Isolan | Fragment, Gesichtet, Iam, SMWFragment, Schutzlevel sysop, Sheikh 2006, Verschleierung |
|
|
|
| Untersuchte Arbeit: Seite: 46, Zeilen: 1-26 |
Quelle: Sheikh 2006 Seite(n): 49, 50, 51, Zeilen: 49: last lines; 50: 11-28 - 51: 1-4 |
|---|---|
| 3.3.3 Enzyme-Linked Immunosorbent Assay (ELISA)
To measure MIP-2/CXCL2 concentration in rat serum, the Quantikine® M rat CXCL2 immunoassay kit (R&D Systems, Wiesbaden, Germany), based on solid phase ELISA, was used. 3.3.3.a. Reagent preparation Since all samples should be pipette within 15 min, reagents needed for the assay were prepared prior to assay procedure. All reagents were provided with Quantikine® immunoassay kit. 3.3.3.b. Rat CXCL2 control The control provided with kit, was reconstituted with 1 ml double distilled water. 3.3.3.c. Rat CXCL2 conjugate concentrate For 96 wells Conjugate concentrate 0.5 ml Conjugate diluent 11 ml 3.3.3.d. Washing buffer For 96 wells Washing buffer concentrate 25 ml ddH2O to 625 ml 3.3.3.e. Substrate solution Equal volumes of color reagents A and B, provided by kit, were mixed together, and solution was used with 15 min. 3.3.3.f. Standard and sample preparation The standards were reconstituted with 2 ml of calibrator diluent. This stock solution (2000 pg/ml) was used to prepare a serial dilution ranging from 31.2 pg/ml to 2000 pg/ml. Calibrator diluent served as zero standard. Prior to assay, serum samples were diluted 2 fold into calibration diluent. |
3.3.1 Enzyme-Linked Immunosorbent Assay (ELISA)
To measure IL-6, IL-1β, TNF-α and IFN-γ concentration in rat serum, the Quantikine® rat IL-6, IL-1β, TNF-α and IFN γ immunoassay kit (R&D Systems, Wiesbaden, Germany), based on solid phase ELISA, was used. [page 50] 3.3.1.II Reagent preparation Since all samples should be pipette within 15 min, reagents needed for the assay were prepared prior to assay procedure. All reagents were provided with Quantikine® immunoassay kit. 3.3.1.III Rat IL-6, IL-1β, TNF-α and IFN-γ control The control provided with kit, was reconstituted with 1 ml double distilled water. 3.3.1.IV Rat IL-6 IL-1β, TNF-α and IFN-γ conjugate concentrate For 96 wells Conjugate concentrate 0.5 ml Conjugate diluent 11 ml 3.3.1.V Washing buffer For 96 wells Washing buffer concentrate 25 ml dd H2O to 625 ml 3.3.1.VI Substrate solution Equal volumes of color reagents A and B, provided by kit, were mixed together, and solution was used with 15 min. 3.3.1.VII Standard and sample preparation [page 51] The standards were reconstituted with 2 ml of calibrator diluent. This stock solution (2000 pg/ml) was used to prepare a serial dilution ranging from 31.2 pg/ml to 2000 pg/ml. Calibrator diluent served as zero standard. Prior to assay, serum samples were diluted 2 fold into calibration diluent. |
The source is not given. |
|
Letzte Bearbeitung dieser Seite: durch Benutzer:Graf Isolan, Zeitstempel: 20140312202228