Rat IL-1 beta/IL-1F2 Quantikine ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.	-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Publication protocol

For enzyme-linked immunosorbent assay (ELISA), rats (n = 18) were randomly divided into three experimental groups (n = 6 per group) as follows: Sham, SCI, and SCI+ high dose β-elemene (320 μg/kg). Segments of the spinal cord (15 mm) including the lesion epicenter at the center were collected 24 h after SCI and homogenized in lysis buffer containing protease inhibitors at 4°C. After 30 min, they were centrifuged at 13, 000 × g at 4°C for 10 min. Then, the protein concentrations of the tissue lysates were measured using a BCA protein assay kit and the lysates were diluted to a final concentration of 1 mg/ml. The ELISA kits for interleukin (IL)-1β (RLB00) and IL-6 (R6000B) were purchased from R&D Systems (Minneapolis, USA). The content of each segment was obtained according to the standard curve at 450 nm minus that at 570 nm.

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Manufacturer protocol

Download the product protocol from R&D Systems for Rat IL-1 beta/IL-1F2 Quantikine ELISA Kit below.

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