Mouse igf 1 quantikine elisa kit

Lysates of whole fetal spleen or liver tissues were prepared using the Qproteome Mammalian Protein Prep Kit (Qiagen, Hilden, Germany), according to the manufacturer's instructions. Hematopoietic cells (CD45+/CD45+Ter119+/Ter119+), endothelial cells (CD45−Ter119−CD31+LYVE-1−), unclassified cells (CD45−Ter119−CD31−LYVE-1−) and CD51+ cells among unclassified cells (CD45−Ter119−CD31−LYVE-1−CD51+) were sorted. Lysates were prepared using M-PER Mammalian Protein Extraction Reagent (Thermo Fisher Scientific, Rockford, IL) and a protease inhibitor from the Qproteome Mammalian Protein Prep Kit (Qiagen). Samples were centrifuged at 12,000 rpm for 10 minutes at 4°C. Supernatants containing soluble protein were collected and protein concentration was quantitated using the Bradford reagent (Bio Rad, Hercules, CA) according to the manufacturer's instruction. The optical density (O.D.) at 540 nm was measured using a Thermo Multiskan FC plate reader (Thermo Fisher Scientific). SCF, IGF-1, IL-3 and EPO ELISA assays were conducted using a mouse SCF Quantikine ELISA kit (R&D Systems, Minneapolis, MN), a mouse IGF-1 Quantikine ELISA kit (R&D Systems), a mouse IL-3 Quantikine ELISA kit (R&D Systems) and a mouse EPO Quantikine ELISA kit (R&D Systems), according to the manufacturer's instructions. O.D.s at 450 nm and 540 nm were measured using a Thermo Multiskan FC plate reader.

Serum IGF1 concentrations in age matched control and irradiated mice were measured using mouse IGF1 Quantikine ELISA kit (Cat# MG100, R&D systems, Minneapolis, MN) as per manufacturer’s instruction. The assay has a detection sensitivity of 8.4 pg/ml.