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One-Step WesternTM Complete Kit (Rabbit)
Kit Description:
Based on GenScript's One-Step WesternTM Blot technology, GenScript One-Step WesternTM Complete Kit reduces the total analysis time for a Western or Dot blot to just one hour. Both chemiluminescent substrates and chromogenic substrates, such as DAB and TMB, are compatible with this kit.
The kit contains WestClearTM Nitrocellulose Membrane (0.2 µm, L00224A60) and LumiSensorTM Chemiluminescent HRP Substrate Kit (L00221V60), optimized for best results. WestClearTM Nitrocellulose Membrane and LumiSensorTM Chemiluminescent HRP Substrate Kit are also available separately.
These kits are not recommended for use with antibodies against phosphoproteins.
Key Features:
Easy to perform: Fewer steps mean fewer chances for human error.
High sensitivity: The kit's sensitivity is comparable to or better than that of the classical 4.5-hour procedure, depending on the quality and amount of antibodies used.
Reproducible results: The kit produces highly reproducible results.
Needs far less optimization than the classical three-step method.
No secondary antibody is needed.
Applications:
The One-Step WesternTM Complete Kit cuts hours off the following applications:
Protein/antigen detection
Evaluation of protein expression
Antibody/antigen titration
Analysis and comparison of primary antibody activity levels
Comparison of One-Step WesternTM Blot detection with classical Western blot detection.
Western blots for the detection of α-Tubulin protein from Hela cell lysate by both classical Western and One-Step WesternTM Complete Kit (Mouse) (L00205C10). 10 µg, 2.5 µg, 0.63 µg, and 0.16 µg of Hela cell lysate (GenScript, M0045) were loaded in different lanes as shown in the figure below. The classical Western blot was developed with ECL system (GE Healthcare, RPN2109). The classical Western blot was developed using the ECL system (GE Healthcare, RPN2109). The One-Step WesternTM Blot was developed using the LumiSensorTM Chemiluminescent HRP Substrate included in the kit.
Western blot detection of housekeeping protein α-Tubulin using polyclonal antibody.
Western blot for the detection of α-Tubulin using the One-Step WesternTM Kit (L00204-C10) and polyclonal α-Tubulin antibody (abcam, ab4074). 10 µg, 5.0 µg, 2.5 µg, 1.25 µg, 0.62 µg, 0.31 µg, 0.16 µg, and 0.08 µg of Hela cell lysate (BD Biosciences, 611449) were loaded into Lane 1, Lane 2, Lane 3, Lane 4, Lane 5, Lane 6, Lane 7, and Lane 8, respectively. The blot was developed with the LumiSensorTM Chemiluminescent HRP Substrate included in the kit.
