One-Step Western^TM Advanced Kit (Rabbit)

Kit Description:

GenScript's Advanced One-Step WesternTM Advanced Kit yields a complete Western blot analysis in just over one hour with even higher sensitivity and lower background than GenScript's standard One-Step WesternTM series. These kits reveal even the tiniest amounts of relevant product. Do not let an inferior Western result overlook small but significant results while absorbing five precious hours of your research time. The One-Step WesternTM Advanced Kit contains all the necessary reagents, buffers, nitrocellulose membrane and HRP substrate for performing a Western blot. It was developed using breakthrough immuno-detection technology (patent pending), allowing the user to perform a complete Western blot in a fraction of the usual time. After transferring the proteins from gel to membrane, simply incubate the membrane in pretreat solution for five minutes and follow with incubation in WB solution (prepared from primary antibody, WB-1 and WB-2) for 40 minutes. Wash the membrane three times for 10 minutes each and then the blot can be developed using the HRP substrate included in the kit. The One-Step WesternTM Advanced Kit procedure is compared to the classical three-step procedure in figure 1 at right. The kit contains WestClearTM Nitrocellulose Membrane (0.2 µm) and LumiSensorTM Plus Chemiluminescent HRP substrate optimized for best results. WestClearTM Nitrocellulose Membrane and LumiSensorTM Plus Chemiluminescent HRP Substrate Kits are also available separately.

Key Features:

• Easy to perform: This kit yields a complete Western analysis in under 90 minutes. No secondary antibody is needed.
• Low background: Faster, surer results with journal-quality images.
• High sensitivity: The sensitivity of this kit is comparable to or higher than that of the classical 4.5-hour procedure, depending on the quality and quantity of antibodies used.

Storage:

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Cat.No. Name Size Price Quantity   L00241 One-Step WesternTM Advanced Kit (Rabbit)  1 Kit (10 Assays) $ 295.00

Technical Manual: Download PDF file TM0223 Components: Components L00241 L00242 L00243 Application  Western detection (rabbit)  Western detection (mouse)  Western detection (goat)  Pretreat A solution  100 ml  100 ml  100 ml  Pretreat B solution  100 ml  100 ml  100 ml  WB-1 solution  1 ml  1 ml  1 ml  WB-2 solution  100 ml  100 ml  100 ml  10X Wash solution  125 ml  125 ml  125 ml  WestClearTM Nitrocellulose Membrane (0.2 µm, 7.5 X 8 cm)  10 Sheets  10 Sheets  10 Sheets  LumiSensorTM Plus Chemiluminescent HRP Substrate  2 x 30 ml  2 x 30 ml  2 x 30 ml  Protocol  1  1  1  EXAMPLE: Zoom PROTOCOL:  20070102153905  (PDF) Examples: Comparison of One-Step WesternTM Adwanced Kit to the classical Western blot using rabbit polyclonal antibody. One-Step WesternTM Advanced Kit is here compared to a classical three-step Western blot for the detection of a His-tagged protein from E. coli cell lysate. Two similar blots were processed with different procedures using Rabbit Anti His-Tag pAb (GenScript, A00174): Classical three-step Western detection (4.5 hours, left panel, Figure 2) and One-Step WesternTM Advanced Kit (1.25 hours, right panel, Figure 2). Figure 2. Western blots for the detection of a His-tagged protein from E. coli cell lysate by both classical Western and One-Step WesternTM using GenScript's One-Step WesternTM Advanced Kit L00241. 200, 80, 32, 12.8, and 5.12 ng of total protein was loaded in Lane 1, 2, 3, 4, and 5, respectively. The classical Western blot was developed with LumiSensorTM Chemiluminescent HRP Substrate (GenScript, L00221V60) and the One-Step WesternTM blot was developed with LumiSensorTM Plus Chemiluminescent HRP Substrate included in the kit L00241. The One-Step WesternTM Advanced Kit produces higher, lower-background signal than the classical three-step procedure while trimming more than four hours off total analysis time.