VALIDATION IMAGES
Human U2-OS cells were stained with Osteocalcin Polyclonal Antibody, Unconjugated(bs-4917R) at 1:200 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
Lane 1: Human MG63 cell lysates; Lane 2: Human A549 cell lysates probed with Osteocalcin Polyclonal Antibody, Unconjugated (bs-4917R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse NIH/3T3 cell lysates; Lane 2: Human MG63 cell lysates; Lane 3: Human A549 cell lysates; Lane 4: Human U-2 OS cell lysates probed with Osteocalcin Polyclonal Antibody, Unconjugated (bs-4917R) at 1:500 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded (human skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Osteocalcin) Polyclonal Antibody, Unconjugated (bs-4917R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Osteocalcin) Polyclonal Antibody, Unconjugated (bs-4917R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Osteocalcin) Polyclonal Antibody, Unconjugated (bs-4917R) at 1:100 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (human skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Osteocalcin) Polyclonal Antibody, Unconjugated (bs-4917R) at 1:100 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-AF488) for 90 minutes, and DAPI for nuclei staining.