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Western Blot
Positive WB detected in: HEK293 whole cell lysate(20µg), Hela whole cell lysate(20µg), A431 whole cell lysate(20µg), Jurkat whole cell lysate(20µg), U-251MG whole cell lysate(20µg), Mouse skeletal muscle tissue lysate(20µg), Rat skeletal muscle tissue lysate(20µg)
All lanes: PKM antibody at 1:1000
Secondary
Goat polyclonal to human IgG at 1/40000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDa
Exposure time: 90s
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IHC image of CSB-RA018072MA1HU diluted at 1:50 and staining in paraffin-embedded human skeletal muscle tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-human polymer IgG labeled by HRP and visualized using 0.05% DAB.
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Immunofluorescence staining of Hela cell with CSB-RA018072MA1HU 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 Fluorescein (FITC) AffiniPure Goat Anti-Human IgG, Fcγ fragment specific.
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Overlay Peak curve showing HepG2 cells stained with CSB-RA018072MA1HU (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 Fluorescein (FITC) AffiniPure Goat Anti-Human IgG, Fcγ fragment specific at 1:200 dilution for 35min at 4℃.Control antibody (green line) was human IgG1 (1ug/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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