HK2 and TK173 cells were stimulated with TGFβ1 (10 ng/ml) ± Tonabersat at 100 μM and 10 μM respectively for 48 h prior to fixing with paraformaldehyde (4%), and subsequent blocking with goat serum (10%) for 1 h at room temperature (RT). Antibodies against fibronectin (Santa Cruz sc‐271098), N‐cadherin (Abcam ab18203), Cx43 (Abcam ab11370), and vimentin (Cell Signaling Technology D21H3), (all 1:200) were used for incubation overnight at 4°C. After washing, cells were incubated with nuclear stain, 4′,6‐diamidino‐2‐phenylindole (DAPI) (1 mmoL/L) for 3 min. Cells were incubated with Alexa‐Fluor 488 for 1 h at RT before visualisation using a Leica TC SP8 confocal microscope (Wetzlar, Germany).
Sc 271098
Manufactured by Abcam
Sc-271098 is a reagent for use in laboratory research. It is a small molecule that can be used as a tool compound. The core function of Sc-271098 is to act as an inhibitor, but no further details on its intended use or target are provided.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700, by Zeiss (4 mentions)
Phase contrast microscope, by Zeiss (2 mentions)
Ab34712, by Abcam (2 mentions)
G2 pro desktop, by Zeiss (2 mentions)
Rhodamine red x, by Thermo Fisher Scientific (2 mentions)
Ab18203, by Abcam (1 mentions)
Tc sp8 confocal microscope, by Leica (1 mentions)
Proteome profiler human cytokine array kit, by R&D Systems (1 mentions)
3 protocols using sc 271098
1
Epithelial-Mesenchymal Transition Markers
2
Characterizing the Surface Morphology and ECM Composition of N-CHDM
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The surface morphology of N-CHDM was observed using phase contrast microscope (Carl Zeiss) and scanning electron microscope (SEM; Phenom G2 Pro Desktop). For immunofluorescence staining of ECM components, N-CHDM was prepared on the glass coverslips, fixed in 4 % paraformaldehyde, then washed with PBS more than three times. Subsequently these samples were treated with 0.2 % Triton X-100 solution for 30 min and they were blocked with 3 % bovine serum albumin (BSA). Once primary antibodies against human fibronectin (Santa Cruz, sc-271,098) and type 2 collagen (COL II) (Abcam, ab34712) were prepared in 1 % bovine serum albumin (BSA) solution, while diluted in 1:200, they were separately treated overnight at 4oC. As the secondary antibodies diluted in 1:500, Alexa fluoro-488 was used for fibronectin and rhodamine red-X (Invitrogen) was applied for COL II. The intensity and distribution of these ECM proteins were confirmed via confocal microscope (LSM700; Carl Zeiss).
3
Characterization of N-CHDM Extracellular Matrix
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The surface morphology of N-CHDM was observed using phase contrast microscope (Carl Zeiss) and scanning electron microscope (SEM; Phenom G2 Pro Desktop). For immunofluorescence staining of ECM components, N-CHDM was prepared on the glass coverslips, fixed in 4% paraformaldehyde, then washed with PBS more than three times. Subsequently these samples were treated with 0.2% Triton X-100 solution for 30 min and they were blocked with 3% bovine serum albumin (BSA). Once primary antibodies against human fibronectin (Santa Cruz, sc-271098) and type 2 collagen (COL II) (Abcam, ab34712) were prepared in 1% bovine serum albumin (BSA) solution, while diluted in 1:200, they were separately treated overnight at 4 o C. As the secondary antibodies diluted in 1:500, Alexa fluoro-488 was used for fibronectin and rhodamine red-X (Invitrogen) was applied for COL II. The intensity and distribution of these ECM proteins were confirmed via confocal microscope (LSM700; Carl Zeiss). The profile of human cytokines within the N-CHDM and S-CHDM were also examined using immunoassay array kit (ARY005B; Proteome Profiler™ Human cytokine Array Kit, R&D system) following the manufacturer's instructions.
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