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ONE-HOUR WesternTM Standard Kit (Rabbit)

ONE-HOUR WesternTM Standard Kit (Rabbit)

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详细说明

GenScript’s ONE-HOUR WesternTM Kit yields journal-quality Western or Dot blot results in about one hour. Using GenScript’s breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step Western process, which can take nearly five hours.

Components:

Components  L00204C  L00204T  L00205T  L00205C  L00228  L00228T 
High-Sensitivity X-Ray Film
(5 x 7 inches) 
20 sheets  20 sheets  20 sheets 
LumiSensorTM Chemiluminescent HRP Substrate  2 x 15 ml  2 x 15 ml  2 x 15 ml 
Pretreat Solution A  100 ml  100 ml  100 ml  100 ml  100 ml  100 ml 
Pretreat Solution B  100 ml  100 ml  100 ml  100 ml  100 ml  100 ml 
Protocol 
WB-1 Solution  1 ml  1 ml  1 ml  1 ml  1 ml  1 ml 
WB-2 Solution  100 ml  100 ml  100 ml  100 ml  100 ml  100 ml 
WestClearTM Nitrocellulose Membrane
(0.2 .m, 7.5 x 8 cm) 
10 sheets  10 sheets  10 sheets  10 sheets  10 sheets  10 sheets 
10X Wash Solution  125 ml  125 ml  125 ml  125 ml  125 ml  125 ml 
ChromoSensorTM One Solution TMB Substrate  60 ml  60 ml  60 ml 


 
 

Kit Description

GenScripts ONE-HOUR WesternTM Kit yields journal-quality Western or Dot blot results in about one hour. Using GenScripts breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step Western process, which can take nearly five hours. Transfer the proteins from gel to membrane and incubate it in the pretreat solution for five minutes. Then incubate in WB-2 solution with the mixture of primary antibody and WB-1 solution for 40 minutes. Then wash three times for five or ten minutes each. The membrane can then be developed with the HRP substrate. The ONE-HOUR WesternTM procedure is contrasted with a classical three-step Western at right.

The ONE-HOUR WesternTM Kit is designed to produce high signal with low background for quick and clear Western analysis of proteins.

Except for basic kits, all other kits contain all the necessary reagents, buffers and materials such as membrane and X-ray films for performing a Western blot, and no additional secondary antibody is needed.

The ONE-HOUR WesternTM Kit saves precious research time. For added cost-efficiency, we recommend our complete kits, which contain all the reagents, membrane, HRP substrate, and X-ray film necessary for a perfect blot. However, the kit components are also sold separately.

Key Features

  • Easy to perform: This kit has fewer and simpler steps than other Western kits.
  • Low background: The kit contains WestClearTM Nitrocellulose Membrane and LumiSensorTM series of Chemiluminescent HRP Substrate Kits, and X-ray film, optimized for low background.
  • Reproducible results: The kit produces highly reproducible results.
  • No additional secondary antibody is needed.

Examples:

Comparison of the two ONE-HOUR WesternTM Blot Kits of different sensitivities using monoclonal antibodies:
Two similar blots were processed with the same procedures using different ONE-HOUR WesternTM Kits: Standard (L00205C) and Advanced (L00242). 10 μg and 2.5 μg of THETM Anti-GST Monoclonal Antibody (Mouse) (GenScript, A00865), respectively, were used with these two kits to detect GST protein.
The results are shown in Figure 1.
ONE-HOUR WesternTM Standard Kit (Rabbit)
Figure 1. Western blots for the detection of GST protein by ONE-HOUR WesternTM using different ONE-HOUR WesternTM Kits: Standard (L00205C) and Advanced (L00242). 33.0, 11.0, 3.60, 1.20, 0.41, 0.13 and 0.045 ng of GST protein were loaded into Lanes 1, 2, 3, 4, 5, 6, and 7 respectively.

Comparison of the two ONE-HOUR WesternTM Blot Kits of different sensitivities using polyclonal antibodies:
Two similar blots were processed with the same procedures using different ONE-HOUR WesternTM Kits: Standard (L00204C) and Advanced (L00241). 10 μg and 2.5 μg of Rabbit Anti-GST-tag Polyclonal Antibody (GenScript, A00097), respectively, were used with the two kits to detect GST protein.
The results are shown in Figure 2.
ONE-HOUR WesternTM Standard Kit (Rabbit)
Figure 2. Western blots for the detection of GST protein by ONE-HOUR WesternTM using different ONE-HOUR WesternTM Kits: Standard (L00204C) and Advanced (L00241). 25.0, 12.5, 6.25, 3.10, 1.50, and 0.78 ng of GST protein were loaded into Lanes 1, 2, 3, 4, 5, and 6 respectively.