The differentiated cells were fixed by being treated with 4% paraformaldehyde for 20 min, at 4 °C. Immunocytochemistry assay was performed on the osteoblast-like cells differentiated from hEnSCs cultured in osteogenic media for 21 days using specific antibodies targeting osteoblast cell markers, such as anti-osteopontin (mouse anti-human, Santa Cruz, USA) and anti-osteocalcin (mouse anti-human, Santa Cruz, USA), and were then incubated with secondary antibody (rabbit anti-mouse IgG-FITC, at a 1:700 dilution; Santa Cruz, USA) for 1 h, at 37 °C as previously described [33 ]. The stained cells were visualized using a fluorescence microscope (Olympus BX51, Japan).
Anti osteocalcin mouse anti human
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-osteocalcin (mouse anti-human) is a laboratory reagent used as a tool for research purposes. It is a monoclonal antibody that specifically binds to the osteocalcin protein, which is a marker of bone formation. The antibody can be used in various research applications, such as Western blotting, immunohistochemistry, and ELISA.
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Lab products found in correlation
2 protocols using anti osteocalcin mouse anti human
1
Osteoblast Differentiation and Characterization
2
Osteocalcin Expression in 3D-Printed Scaffolds
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The immunocytochemistry
(ICC)
technique was performed to detect the expression levels of osteocalcin
on the 3D-printed scaffolds. For this purpose, after being in osteogenic
culture for 21 days, the cell suspension was cultured on sterile gelatin
lamellae. After 24 h, it was washed with PBS and fixed at 4 °C
for 20 min with 4% paraformaldehyde. Lamellae were incubated in 2
N HCL for 20 min at room temperature, after washing with PBS. Next,
the lamellae were exposed to 0.3% Triton X-100 for 30 min (Triton-permeable
cell membrane to antibodies). Afterward, 10% goat serum was added
to the cells for half an hour (goat serum proteins cause nonspecific
antigen sites to be coated and prevent a nonspecific reaction). Cells
were then incubated overnight with primary antibodies; anti-osteocalcin
(mouse anti-human, Santa Cruz) at a 1:100 (PBS) dilution at 4 °C.
They were then washed twice with PBS and exposed to conjugated secondary
antibodies (rabbit anti-mouse IgG-FITC, Santa Cruz) for 60 min at
a 1:200 (PBS) dilution and 37 °C in the dark. After three times
washing with PBS, DAPI was used to stain the nuclei and then examined
with a fluorescence microscope (LABOMED).
(ICC)
technique was performed to detect the expression levels of osteocalcin
on the 3D-printed scaffolds. For this purpose, after being in osteogenic
culture for 21 days, the cell suspension was cultured on sterile gelatin
lamellae. After 24 h, it was washed with PBS and fixed at 4 °C
for 20 min with 4% paraformaldehyde. Lamellae were incubated in 2
N HCL for 20 min at room temperature, after washing with PBS. Next,
the lamellae were exposed to 0.3% Triton X-100 for 30 min (Triton-permeable
cell membrane to antibodies). Afterward, 10% goat serum was added
to the cells for half an hour (goat serum proteins cause nonspecific
antigen sites to be coated and prevent a nonspecific reaction). Cells
were then incubated overnight with primary antibodies; anti-osteocalcin
(mouse anti-human, Santa Cruz) at a 1:100 (PBS) dilution at 4 °C.
They were then washed twice with PBS and exposed to conjugated secondary
antibodies (rabbit anti-mouse IgG-FITC, Santa Cruz) for 60 min at
a 1:200 (PBS) dilution and 37 °C in the dark. After three times
washing with PBS, DAPI was used to stain the nuclei and then examined
with a fluorescence microscope (LABOMED).
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