Trident femto Western HRP Substrate

293T whole cell extracts were separated by 10% SDS-PAGE, and the membrane was blotted with alpha Tubulin antibody [HL2162] (GTX638140) diluted at 1:2000 and then with goat anti-rabbit HRP-conjugated secondary antibody (GTX213110-01). Identical blots were incubated with 1 mL of Trident femto Western HRP Substrate (GTX14698), SuperSignal™ West Femto Maximum Sensitivity Substrate, and ECL™ Prime Western Blotting System. The blots were simultaneously exposed for 9 seconds and 1 seconds using Fusion SOLO 6S EDGE imaging system. SuperSignal™ West Femto, ECL™ are registered trademark of Thermo Fisher Scientific and Cytiva, respectively. The trademark holder is not affiliated with GeneTex and does not endorse this product.

Recombinant protein mCherry was loaded 100, 20, 4 and 0.8 pg and the blot was probed with mCherry antibody (GTX128508) diluted at 1:500. A HRP-conjugated secondary antibody was applied and developed with 1 ml of Trident femto Western HRP Substrate (GTX14698). Exposure time : 18 and 54 seconds. Immobilon is a registered trademark which is not affiliated with GeneTex and does not endorse this product.

Serial dilutions of purified mCherry recombinant protein were prepared and applied at 10 μL/well. After electrophoresis and protein transfer, membranes were probed with anti-mCherry antibody (GTX128508) diluted at 1:1500 and then with goat anti-rabbit HRP-conjugated secondary antibody (1:2000). Identical blots were incubated with 1 ml of Trident femto Western HRP Substrate, SuperSignalTM West Femto, Trident pico Western HRP Substrate, ClarityTM, and LuminataTM Forte. The blots were simultaneously exposed for 90 seconds and 180 seconds using BioSpectrumTM 500 digital imaging system SuperSignalTM West Femto, ClarityTM, LuminataTM Forte are registered trademark of Thermo Fisher Scientific, Bio-Rad, and EMD Millipore, respectively. The trademark holder is not affiliated with GeneTex and does not endorse this product.

Recombinant protein mcherry is loaded as the follows: Lane A 3.13 ng recombinant protein Lane B 1.56 ng recombinant protein Lane C 780 pg recombinant protein Lane D 390 pg recombinant protein The blot was probed with mcherry antibody (GTX128508) diluted at 1:2000. A HRP-conjugated secondary antibody was applied and developed with 1 ml of Trident femto Western HRP Substrate (GTX14698). Exposure time : 18 seconds, 54 seconds, and 90 seconds. SuperSignalTM is a registered trademark of Thermo Fisher Scientific. The trademark holder is not affiliated with GeneTex and does not endorse this product.

Various 293T whole cell extracts were prepared and applied at 10 μL/well. After electrophoresis and protein transfer, membranes were probed with alpha Tubulin antibody (GTX112141) diluted at 1:100000 and then with goat anti-rabbit HRP-conjugated secondary antibody (GTX213110-01). Identical blots were incubated with 1 ml of Trident femto Western HRP Substrate and the mixed substrate (Substrate solution A and Substrate solution B) stored for 0, 15 and 24 hours at 2-8ºC. The blots were simultaneously exposed for 18 seconds, 90 seconds and 180 seconds using BioSpectrumTM 500 digital imaging system. The signal was developed with Trident femto Western HRP Substrate (GTX14698).