Mouse CXCL12/SDF-1 alpha 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

Since CCL5 is a relatively well-characterised component of the innate immune response, but is not represented on the Luminex assay employed for the other cytokines/chemokines, CCL5 levels in maternal plasma and embryonic brain tissue supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Additional measurements of CXCL10 and CXCL12 levels in placenta were also performed by ELISA. Mouse Quantikine® ELISAs, MMR00, MCX100 and MCX120 (R&D Systems, Abingdon, UK), were utilised according to the manufacturer’s instructions. Fifty microlitre of assay diluent (provided) and 50 μl of the diluted standards and samples (undiluted) were assayed in duplicate. The optical density was then read by a plate reader (Multiskan Spectrum, Thermo Fisher). Embryonic brain and placenta concentrations were normalised to total protein as established by the BCA assay.

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse CXCL12/SDF-1 alpha Quantikine ELISA Kit below.

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