Rat Bone Morphogenetic Protein 2 ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Avoid repeated freeze-thaw cycles. -Prior to assay, the frozen sample should be brought to room temperature slowly and mixed gently. -Do not use heat-treated specimens.	-Bring all kit components and samples to room temperature (20-25 °C) before use. -Avoid sunlight for Substrate Solution. -Read absorbance immediately after adding the stop solution. -TMB is light sensitive, avoid prolonged exposure to light. Also avoid contact between TMB and metal, otherwise color may develop.

Upstream tips
- Avoid repeated freeze-thaw cycles. -Prior to assay, the frozen sample should be brought to room temperature slowly and mixed gently. -Do not use heat-treated specimens.
Protocol tips
-Bring all kit components and samples to room temperature (20-25 °C) before use. -Avoid sunlight for Substrate Solution. -Read absorbance immediately after adding the stop solution. -TMB is light sensitive, avoid prolonged exposure to light. Also avoid contact between TMB and metal, otherwise color may develop.

Publication protocol

The morning fasting peripheral venous blood (5 ml) was collected on the next day from all patients who were confirmed to have osteoporotic fracture on admission and centrifuged at 2000 rpm for 10 min, followed by serum separation and storage at –80°C. After experiments, the rats were anesthetized by 3% pentobarbital sodium, which was followed by transabdominal collection of arterial blood (4 ml), centrifugation at 4000 rpm for 10 min, serum separation, and storage at –20°C. Serum levels of BMP2 and Smad4 and the activity of alkaline phosphatase (ALP) in patients and rats were detected using the double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit (Blue Gene). These serum samples (100 μl) were collected, followed by reconstitution at room temperature, re-centrifugation, and collection of supernatant for measurement. The optic density (OD) value was measured using a microplate reader at 450 nm.

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Manufacturer protocol

Download the product protocol from Blue Gene for Rat Bone Morphogenetic Protein 2 ELISA below.

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