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Primaries antibodies (prediluted in blocking solution) were incubated ON at 4°C (Rabbit anti HA 1∶300, Bethyl Cat. No. A190-108A; Mouse anti Synaptopodin 1∶80, Progen Cat. No. 61094).
Primary antibodies diluted in blocking solution were incubated over night at 4 °C.
Primary antibodies prepared in blocking solution were incubated overnight at 4ºC.
pY397FAK, vinculin, or pY118 paxillin antibodies (1 100 in blocking solution) were incubated overnight at 4°C.
Different dilutions (1 10,000 to 1 2,000) of rabbit primary polyclonal antibodies (H3K9me3 antibody ab8898 lot GR102573-1, H3K4me3 antibody ab8580 lot GR56122-1) in blocking solution were incubated with the array under mild rocking at 4°C for 1 hour.
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100 μL of primary anti-elongin B mAb diluted (1 500) in blocking solution was incubated overnight at 4 °C.
A total of 250 µL scFv-Fc fusion protein with a final concentration of 2 µg/mL in blocking solution was incubated for 1.5 h and washed three times with PBS.
Coverslips were washed and incubated in 1X PBS pH 7.4 containing 5% FBS and 3% BSA (blocking solution) for at least 30 min. Primary antibody diluted in blocking solution was incubated for 30 60 min, the coverslips washed 3X with blocking buffer and then incubated for 30 60 min with secondary antibodies: Alexa Fluor 594 for mouse diluted 1 2000 in blocking buffer.
Primary antibody (in blocking solution) was incubated for 60 min and then washed three times with blocking solution.
Horseradish peroxidase-conjugated secondary antibody (1/5000 or 1/1000 [for cyclin D1] dilution) in blocking solution was incubated with the membrane for 1 h at 20°C.
An anti-Sjda monoclonal antibody (2 mg/ml) diluted 1 500 in blocking solution was incubated with the parasites at 4°C overnight.
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