F-actin staining was performed according to the manufacturer’s instructions (Invitrogen#R415). Briefly, 1.5 × 105 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1( +)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 μL containing 4 μL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC-conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immunofluorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
Sc 365846 fitc
Manufactured by Santa Cruz Biotechnology
Sc-365846 FITC is a fluorescently-labeled antibody product manufactured by Santa Cruz Biotechnology. It is designed for use in various laboratory techniques that require fluorescent detection.
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Lab products found in correlation
5 protocols using sc 365846 fitc
1
Cytoskeleton Visualization and Protein Localization
2
Investigating ARHGAP5-AS1 Translation and Actin Dynamics
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This experiment was performed according to the manufacturer's instructions (Promega#L1170). Brie y, 1 μg of circular plasmid pcDNA3.1(+)-ARHGAP5-AS1 and pcDNA3.1(+)-ARHGAP5-AS1-AS were used in the translation reaction. The tubes were incubated 1 h at 30 ℃. Then, 2 μL of reaction liquid was added into 15 μL SDS Loading Buffer and boiled 3 times at 105 ℃ on incubator. Streptavidin-HRP (CST#3999) was utilized to detect products of translation.
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 μL containing 4 μL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 μL containing 4 μL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
3
Investigating ARHGAP5-AS1 Translation and Actin Dynamics
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This experiment was performed according to the manufacturer's instructions (Promega#L1170). Brie y, 1 μg of circular plasmid pcDNA3.1(+)-ARHGAP5-AS1 and pcDNA3.1(+)-ARHGAP5-AS1-AS were used in the translation reaction. The tubes were incubated 1 h at 30 ℃. Then, 2 μL of reaction liquid was added into 15 μL SDS Loading Buffer and boiled 3 times at 105 ℃ on incubator. Streptavidin-HRP (CST#3999) was utilized to detect products of translation.
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 μL containing 4 μL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 μL containing 4 μL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
4
In Vitro Translation and Immunofluorescence Analysis of ARHGAP5-AS1
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This experiment was performed according to the manufacturer's instructions (Promega#L1170). Brie y, 1 µg of circular plasmid pcDNA3.1(+)-ARHGAP5-AS1 and pcDNA3.1(+)-ARHGAP5-AS1-AS were used in the translation reaction. The tubes were incubated 1 h at 30 ℃. Then, 2 µL of reaction liquid was added into 15 µL SDS Loading Buffer and boiled 3 times at 105 ℃ on incubator. Streptavidin-HRP (CST#3999) was utilized to detect products of translation.
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 µL containing 4 µL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
Immuno uroscence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 µL containing 4 µL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
5
Protein Expression and Cytoskeleton Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This experiment was performed according to the manufacturer's instructions (Promega#L1170). Brie y, 1 µg of circular plasmid pcDNA3.1(+)-ARHGAP5-AS1 and pcDNA3.1(+)-ARHGAP5-AS1-AS were used in the translation reaction. The tubes were incubated 1 h at 30 ℃. Then, 2 µL of reaction liquid was added into 15 µL SDS Loading Buffer and boiled 3 times at 105 ℃ on incubator. Streptavidin-HRP (CST#3999) was utilized to detect products of translation.
Immuno uorescence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 µL containing 4 µL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
Immuno uorescence staining F-actin staining was performed according to the manufacturer's instructions (Invitrogen#R415). Brie y, 1.5 × 10 5 cells transfected with si-NC/si-ARHGAP5-AS1 or pcDNA3.1(+)-VEC/pcDNA3.1(+)-ARHGAP50AS1 for 48 h were seeded onto coverslips in 24-well culture plate. A total volume of 200 µL containing 4 µL Rhodamine Phalloidin was used per well and incubated 30 min at room temperature. As for SMAD7 staining, the FITC conjugated antibody (Santa Cruz#sc-365846 FITC) was used at the dilution rate of 1:50 in 1% BSA and incubated overnight. Immuno uorescence pictures were taken by Nikon A1R confocal microscope (Nikon, Kanagawa, Japan).
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