SW480 cell lines were seeded on coverslips and cultured in dishes at 37 °C with 5% CO2, when 80% confluent cells were formed. 20 μM recombinant antibody RGD4C-scFv was added, and incubated for 5 h at 37 °C. And then fixed with 4% paraformaldehyde for 30 min. After permeabilized with PBS containing 0.2% Triton X-100 (Sigma-Aldrich, Darmstadt, Germany) and washed with PBS containing 0.02% Tween-20 (PBST)three-times, the slides were incubated overnight at 4 °C with primary rabbit anti-His Tag mAb (clone number: D3I1O, Cell Signaling TECHNOLOGY, USA) and mouse pan-Ras mAb (clone number: C4, SANTA CRUZ, USA), washed for 5 min with PBS then incubated for 1 h at 37 °C in the dark with FITC-conjugated goat anti-rabbit antibody (ZSGB-BIO) and TRITC-conjugated goat anti-mouse antibody (ZSGB-BIO). Nuclei were stained with 4′,6-diamidino-2-phenylindole (Sigma, Da, Germany) at 25 °C for approximately 15–20 min. The fluorescence signals were analyzed with a fluorescence microscope (OlympusBX51, Tokyo, Japan).
Mouse pan ras mab
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Mouse pan-Ras mAb is a monoclonal antibody that recognizes all Ras isoforms (H-Ras, K-Ras, and N-Ras). It can be used for the detection of Ras proteins in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Primary rabbit anti his tag mab, by Cell Signaling Technology (2 mentions)
Fitc conjugated goat anti rabbit antibody, by Santa Cruz Biotechnology (1 mentions)
Fitc conjugated goat anti rabbit antibody, by ZSGB-BIO (1 mentions)
3 protocols using mouse pan ras mab
1
Visualization of RGD4C-scFv Binding
2
Visualizing Ras Protein Interaction
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The cell lines were seeded on coverslips and cultured in dishes at 37°C with 5% CO2, when 80% confluent cells were formed. BR2-scFv was added, and cocultured for 7 hours. And then fixed with 4% paraformaldehyde for 30 min and permeabilized with 0.2% Triton 100 for 20 minutes.The slides were incubated overnight at 4°C with primary rabbit anti-His Tag mAb (clone number: D3I1O, Cell Signaling TECHNOLOGY, USA) and mouse pan-Ras mAb (clone number: C4, SANTA CRUZ, USA), washed for 5 min with PBS then incubated for 1h at 37°C in the dark with FITC-conjugated goat anti-rabbit antibody (ZSGB-BIO) and TRITC-conjugated goat anti-mouse antibody (ZSGB-BIO). The nuclei were stained with DAPI, and the results were observed under a fluorescence microscope.
3
Visualizing Antibody Binding in SW480 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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SW480 cell lines were seeded on coverslips and cultured in dishes at 37 °C with 5% CO 2 , when 80% confluent cells were formed. 20 μM recombinant antibody RGD4C-scFv was added, and incubated for 5 h at 37 °C. And then fixed with 4% paraformaldehyde for 30 min. After permeabilized with PBS containing 0.2% Triton X-100 (Sigma-Aldrich, Darmstadt, Germany) and washed with PBS containing 0.02% Tween-20 (PBST)threetimes, the slides were incubated overnight at 4 °C with primary rabbit anti-His Tag mAb (clone number: D3I1O, Cell Signaling TECHNOLOGY, USA) and mouse pan-Ras mAb (clone number: C4, SANTA CRUZ, USA), washed for 5 min with PBS then incubated for 1 h at 37 °C in the dark with FITC-conjugated goat antirabbit antibody (ZSGB-BIO) and TRITC-conjugated goat anti-mouse antibody (ZSGB-BIO). Nuclei were stained with 4′,6-diamidino-2-phenylindole (Sigma, Da, Germany) at 25 °C for approximately 15-20 min. The fluorescence signals were analyzed with a fluorescence microscope (OlympusBX51, Tokyo, Japan).
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