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[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

Typus
Verschleierung
Bearbeiter
Hindemith
Gesichtet
Yes.png
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.

Anmerkungen

The source is not given.

Sichter
(Hindemith) Schumann


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