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Western Blot
Positive WB detected in: 25ng Recombinant protein with GST tag from E.coli
All lanes: GST antibody at 1:10000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 32.6, 30.1, 29.5, 37.4, 30.4, 60.5, 38.1, 61.6 KDa
Observed band size: 31, 30, 31, 37, 30, 60, 37, 61 KDa
Exposure time：5min
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Western Blot
Positive WB detected in: 50ng Recombinant protein with GST tag from E.coli
All lanes: GST antibody at 1:2000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 26, 102.9, 75.6 KDa
Observed band size: 26, 103, 76 KDa
Exposure time：1min
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Western Blot
Positive WB detected in: Recombinant protein with GST tag from E.coli at 50ng, 25ng, 12.5ng, 6.25ng, 3.125ng, 1.5625ng
All lanes: GST antibody at 1:10000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 26 KDa
Observed band size: 26 KDa
Exposure time：5min
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Western Blot
Positive WB detected in: 25ng recombinant protein with GST tag from E.coli
GST antibody at 1:10000, 1:20000, 1:40000, 1:80000, 1:160000, 1:320000, 1:640000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 26 KDa
Observed band size: 26 KDa
Exposure time：5min
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Immunofluorescence staining of 293F transfected cells with CSB-MA000031M1m at 1:94.5, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
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Immunoprecipitating GST in transfected-293F whole cell lysate
Lane 1: Mouse control IgG instead of CSB-MA000031M1m in 293F cell lysate Transfected with GST
Lane 2: CSB-MA000031M1m (5µg) + Transfected-293F whole cell lysate (500µg)
Lane 3: Transfected-293F whole cell lysate (10µg)
For western blotting, the blot was detected with CSB-MA000031M1m at 1:2000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:50000
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Overlay Peak curve showing 293F cells transfected with GST stained with CSB-MA000031M1m (red line) at 1:189. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG2b (1µg/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
