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Western Blot
Positive WB detected in: A549 whole cell lysate(30µg), U251 whole cell lysate(30µg),hela whole cell lysate(30µg), U87 whole cell lysate(30µg), BXPC-3 whole cell lysate(30µg), HEK293 whole cell lysate(30µg), PC-3 whole cell lysate(30µg)
All lanes: PKM antibody at 1:1000
Secondary
Goat polyclonal to Anti-Mouse IgG (H+L) at 1/40000 dilution
Predicted band size: 57.937, 58.062, 56.273 kDa
Observed band size: 58 kDa
Exposure time: 15s
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Immunofluorescence staining of A-549 cell with CSB-RA018072MA2HU at 1:30 ,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was FITC-conjugated Goat Anti-Mouse IgG(H+L).
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Immunofluorescence staining of U-251MG cell with CSB-RA018072MA2HU at 1:30 ,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was FITC-conjugated Goat Anti-Mouse IgG(H+L).
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Overlay Peak curve showing HepG2 cells stained with CSB-RA018072MA2HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*106cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1:200 dilution for 35min at 4℃.Control antibody (green line) was mouse IgG (1ug/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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