Western blot analysis of GADD153/DDIT3 using anti-GADD153/DDIT3 antibody (A00311-4). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: RAW264.7 whole cell lysates,
Lane 2: C6 whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GADD153/DDIT3 antigen affinity purified polyclonal antibody (A00311-4) at a dilution of 1:1000 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for GADD153/DDIT3 at approximately 29 kDa. The expected band size for GADD153/DDIT3 is at 19 kDa.
Flow Cytometry analysis of C6 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing C6 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of Raw264.7 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing Raw264.7 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
| Western blot (WB): | 1:500-2000 |
| Flow Cytometry (Fixed): | 1:50-200 |
| Enzyme linked immunosorbent assay (ELISA): | 1:100-1000 |
Western blot analysis of GADD153/DDIT3 using anti-GADD153/DDIT3 antibody (A00311-4). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: RAW264.7 whole cell lysates,
Lane 2: C6 whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GADD153/DDIT3 antigen affinity purified polyclonal antibody (A00311-4) at a dilution of 1:1000 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for GADD153/DDIT3 at approximately 29 kDa. The expected band size for GADD153/DDIT3 is at 19 kDa.
Flow Cytometry analysis of C6 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing C6 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of Raw264.7 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing Raw264.7 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Western blot analysis of GADD153/DDIT3 using anti-GADD153/DDIT3 antibody (A00311-4). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: RAW264.7 whole cell lysates,
Lane 2: C6 whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GADD153/DDIT3 antigen affinity purified polyclonal antibody (A00311-4) at a dilution of 1:1000 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for GADD153/DDIT3 at approximately 29 kDa. The expected band size for GADD153/DDIT3 is at 19 kDa.
Flow Cytometry analysis of C6 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing C6 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of Raw264.7 cells using anti-GADD153/DDIT3 antibody (A00311-4).
Overlay histogram showing Raw264.7 cells stained with A00311-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GADD153/DDIT3 Antibody (A00311-4) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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