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Separated proteins were electro-transferred onto a nitrocellulose membrane by iBlot™ Gel Transfer Device (Invitrogen), and the membrane was cut into strips corresponding to sample load of 5 salivary glands per strip.
For Western blot, samples were loaded onto a NuPAGE® Novex® 4 12% Bis-Tris Gel (Life Technologies) and blotted using in an iBlot Gel Transfer Device (Life Technologies).
Cell lysates were electrophoresed in 4 15% Mini-PROTEAN® TGX™ Precast Protein Gels (Bio-Rad), transferred to PVDF membranes by iBlot™ Gel Transfer Device (ThermoFisher), and blotted with the corresponding primary and secondary antibodies.
Equal amount of proteins (20 μg/sample) were loaded and separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto polyvinylidene difluoride membranes using iBlotTM2 gel transfer system (Invitrogen).
The DNA/protein mixtures were then analyzed by the native 7% PAGE, and directly transferred onto nylon membrane by contact blotting-aided gel transfer.
Through the process of making family photographs digital, printing them, creating a gel transfer on my body, and creating a photograph, which was then silver printed, each step removes me from the original.
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Supernatants were separated on a 10% SDS-PAGE gel, transferred to polyvinylidene fluoride membrane.
Total proteins were separated on a 12% SDS/PAGE gel, transferred to nitrocellulose membrane (Whatman).
Samples were analyzed by SDS-polyacrylamide gel, transferred to nitrocellulose membranes and autoradiographed.
For immunoblotting, protein samples were separated on a 6% SDS-PAGE gel, transferred to Immobilon™ Transfer Membrane (Millipore, Bedford, MA).
Gel transfers were performed in a Hoeffer wire transfer tank at 50 volts for 2 hours at 4°C.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com