Immunocytochemical staining was performed in vitro to characterize human primary tenocytes following a standard protocol [11 ], using antibodies against COL1 (1:200), COL3 (1:200), vimentin (1:200), SCX (1:400), CD34 (1:200), and OCT4 (1:200). When using mouse primary antibodies, normal goat serum (Solarbio; A8020) was used for blocking and a goat anti-mouse secondary antibody conjugated with fluorescein isothiocyanate (FITC) (Bioss; bs-0296G-FITC) was applied. For rabbit primary antibodies, normal goat serum (Solarbio) and a goat anti-rabbit secondary antibody (Bioss; bs-0295G-AF594) were used. Cell nuclei were counterstained with DAPI-containing mounting media (Solarbio; S2110) and analyzed under a fluorescence microscope (Olympus BX53; Olympus Corporation, Tokyo, Japan). The montages were created using Adobe Photoshop (version CC2017; Adobe, San Jose, CA, USA).
Bs 0295g af594
Manufactured by Bioss Antibodies
The Bs-0295G-AF594 is a fluorescently labeled antibody produced by Bioss Antibodies. It is designed for use in various research applications that require the detection and visualization of specific target molecules or cellular structures.
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2 protocols using bs 0295g af594
1
Immunocytochemical Characterization of Human Tenocytes
2
Detecting Phospho-PERK in HOS Cells
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Following treatment, processed HOS cells were stabilized using 4% paraformaldehyde for a duration of 20 min, made permeable with 0.1% Triton-X 100 for 5 min, and then blocked using 3% bovine serum albumin (BSA) and 0.08% glycine for a period of 50 min. Samples were incubated with polyclonal rabbit anti-phospho-PERK antibody (bs-3330R, Bioss, USA) at 1:150 overnight at 4°C and then with FITC-conjugated goat anti-rabbit secondary antibody (bs-0295G-FITC, Bioss) at 1:200 for 1 h. For sections, paraffin-embedded slides were prepared as described in section 2.14 (Histology). Sections were deparaffinized, hydrated and subjected to antigen retrieval as described previously. For immunofluorescence (IF) examination of tissues, sections were also permeabilized and blocked before incubation with anti-phospho-PERK primary antibody and goat anti-rabbit Fluor 594 secondary antibody (bs-0295G-AF594, Bioss). Then, nuclei were visualized with DAPI (Beyotime Biotech) staining, and samples were imaged by fluorescence microscopy. Fluorescence intensity was analyzed using ImageJ 1.52.
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