Anti pgrn polyclonal antibody

Seven IVD samples from patients with disc degeneration were harvested in this study, and Institutional Review Boards (IRB#2852 from Sutter Medical Center in California) approved the experiments. Informed consent was obtained from all subjects. Besides, IVD tissue from 4-, 6- and 9-month old WT mice were harvested and fixed in 4% PBS buffered paraformaldehyde at 4°C overnight. After the tissue was dehydrated and embedded in paraffin, 6-μm sections were cut. Thereafter, sections were deparaffinized by xylene immersion, rehydrated by graded ethanol, and treated with 0.1% trypsin for 30 minutes at 37°C. After blocking in 20% goat serum for 60 minutes at room temperature, sections from human IVD were incubated with anti-PGRN polyclonal antibody (1:100 dilution; Santa Cruz Biotechnology), and sections from indicated mice were incubated with anti-neoepitope of aggrecan (1:100 dilution; Millipore, Cat. No: AB8135), anti-phosphorylated IκB-α (pIκB-α) (1:100 dilution; Santa Cruz Biotechnology; Cat. No. SC-101713) or anti-β-catenin polyclonal antibody (1:100 dilution; Santa Cruz Biotechnology; Cat. No. SC-1496) at 4°C overnight, followed by incubation with a horseradish peroxidase–conjugated secondary antibody for 60 minutes at room temperature. The signal was detected using the Vector Elite ABC Kit (Vectastain; Vector).

Seventeen IVD samples from patients with disc degeneration were harvested with approval of Institutional Review Boards (IRB#2852 from Sutter Medical Center in California). Besides, IVD tissue from 2-, 4-, 6- and 9-month old WT mice were harvested and fixed in 4% PBS buffered paraformaldehyde at 4°C overnight for immunohistochemistry. After the tissue was dehydrated and embedded in paraffin, 6-μm sections were cut. Thereafter, sections were deparaffinized by xylene immersion, rehydrated by graded ethanol, and treated with 0.1% trypsin for 30 minutes at 37°C. After blocking in 20% goat serum for 60 minutes at room temperature, sections from human IVD were incubated with anti-PGRN polyclonal antibody (1:100 dilution; Santa Cruz Biotechnology), and sections from 6-month old mice were incubated with anti-neoepitope of aggrecan (1:100 dilution;Millipore, Cat. No: AB8135), anti-phosphorylated IκB-α (pIκB-α) (1:100 dilution; Santa Cruz Biotechnology; Cat. No. SC-101713) or anti-β-catenin polyclonal antibody (1:100 dilution; Santa Cruz Biotechnology; Cat. No. SC-1496) at 4°C overnight, followed by incubation with a horseradish peroxidase–conjugated secondary antibody for 60 minutes at room temperature. The signal was detected using the Vector Elite ABC Kit (Vectastain; Vector).