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ONE-HOUR Western^TM Complete Kit Using TMB (Rabbit)

Kit Description

GenScript's ONE-HOUR Western^TM 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. 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 Western^TM procedure is contrasted with a classical three-step Western at right.

The ONE-HOUR Western^TM 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 Western^TM 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 WestClear^TM Nitrocellulose Membrane and LumiSensor^TM 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.

Order:

Cat.No. Name Size Price Quantity   L00204T ONE-HOUR WesternTM Complete Kit Using TMB (Rabbit)  1 Kit (10 Assays) $ 255.00

Technical Manual: Download PDF file TM0352 Components: Components L00228 L00204C L00204T L00205C L00205T L00228T 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  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  10X Wash Solution  125 ml  125 ml  125 ml  125 ml  125 ml  125 ml  WestClearTM Nitrocellulose Membrane (0.2 µm, 7.5 x 8 cm)  10 sheets  10 sheets  10 sheets  10 sheets  10 sheets  10 sheets  ChromoSensorTM One Solution TMB Substrate  X  X  60 ml  X  60 ml  60 ml  LumiSensorTM Chemiluminescent HRP Substrate  2 x 15 ml  2 x 15 ml  X  2 x 15 ml  X  X  High-Sensitivity X-Ray Film (5 x 7 inches)  20 sheets  20 sheets  X  20 sheets  X  X  Protocol  1  1  1  1  1  1  MSDS:  20060818160706  (PDF) Reference: Zhang W, et al. Transcriptional down-regulation of thromboxane A(2) receptor expression via activation of MAPK ERK1/2, p38/NF-kappaB pathways. J. Vasc. Res. 2009;46(2):162-74. 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. Figure 1. Western blots for the detection of GST protein by ONE-HOUR WesternTM using different One-Step 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-Step WesternTM Blot Kits of different sensitivities using polyclonal antibodies: Two similar blots were processed with the same procedures using different One-Step 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. Figure 2. Western blots for the detection of GST protein by One-Step WesternTM using different One-Step 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.

Related Products:   Code Name Size Price Quantity L00204  ONE-HOUR WesternTM Basic Kit (Rabbit)  1 Kit (10 Assays)  $ 195.00 L00205  ONE-HOUR WesternTM Basic Kit (Mouse)  1 Kit (10 Assays)  $ 195.00 L00224A60  WestClearTM Nitrocellulose Membrane  1 pk (10 sheets of 7.5 x 8 cm)  $ 60.50 L00221V60  LumiSensorTM Chemiluminescent HRP Substrate Kit  1 Kit (2 X 30ml)  $ 55.00 L00222V60  ChromoSensorTM One-Solution TMB Substrate  1 pk (60ml)  $ 28.00 L00205T  ONE-HOUR WesternTM Complete Kit Using TMB (Mouse)  1 Kit (10 Assays)  $ 255.00 L00228  ONE-HOUR WesternTM Complete Kit (Goat)  5 kit (50 assays)  $ 962.50 L00228  ONE-HOUR WesternTM Complete Kit (Goat)  1 Kit (10 Assays)  $ 275.00 L00228T  ONE-HOUR WesternTM Complete Kit Using TMB (Goat)  1 kit  $ 255.00 L00354  LumiSensorTM Super HRP Substrate  1 kit  $ 135.00 L00358  High-Sensitivity X-Ray Film (5 X 7 inches)  100 sheets  $ 65.00 L00358  High-Sensitivity X-Ray Film (5 X 7 inches)  20 sheets   $ 14.90