Goat bmp 2 primary antibody

The tissue sections were washed twice in 0.1 M phosphate-buffered saline (PBS), twice in 1% triton X-100 (Sigma) for 15 min, and then twice with 0.5% bovine serum albumin (BSA; Sigma) dissolved in PBS for 15 min. The sections were then incubated with goat BMP-2 primary antibody and rabbit IGF-1 primary antibody (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at room temperature in a humidity chamber. After 24 h, the sections were washed twice with 0.5% BSA in PBS and then incubated with either biotinylated anti-goat secondary antibody (1:200; Vector Laboratories, Burlingame, CA, USA) or biotinylated anti-rabbit secondary antibody (1:200; Jackson Immuno Research Laboratories, West Grove, PA, USA) for 1 h. After being washed twice with PBS for 15 min, the sections were incubated with an avidin-biotin-peroxidase complex (1:100, Vectastain ABC Kit; Vector Laboratories) for 1 h at room temperature. After another wash with PBS, the sections were stained and reacted with a 0.05% 3, 3-diaminobenzidine (DAB) solution containing hydrogen peroxide in PBS. The reaction was stopped by washing them with PBS and then the slides were dehydrated with solutions of 50, 75, 95, and 100% ethanol and xylene, in that order. The sections were mounted on glass slides with Permount medium solution (Fisher Scientific, Waltham, MA, USA) and micrographs of the sections were taken.

After pretreatment as previously described [15 (link)], the sagittal sections were incubated overnight with rabbit IGF1 and goat BMP2 primary-antibody (1/200, Santa Cruz, Dallas, TX, USA), respectively. After washing, the sections were incubated for an hour with rabbit secondary-antibody (1/200, Jackson, West Grove, PA, USA). After washing, sections were incubated with avidin-biotin complex reagent (1/100, Vector, USA) for 1 h. Sections were stained with a 0.05% 3,3-diaminobenzidine (Sigma) with hydrogen peroxide.

The sagittal parts were incubated overnight at 4 °C with goat BMP-2 primary-antibody and rabbit IGF-1 (1:200, Santa Cruz, Dallas, TX, USA) after pretreatment [19 (link)]. After cleaning, the parts were incubated with rabbit secondary-antibody (1:200, Jackson, West Grove, PA, USA) for 1 h. Subsequent to washing, parts were incubated with avidin-biotin complex reagent (1:100, Vector, Burlingame, CA, USA) for an hour. The parts were stained with a 3,3-diaminobenzidine (0.05%) and hydrogen peroxide.