InstantBlot - 1 Kit

InstantBlot - 1 Kit

Development of western blots with secondary antibodies while simple in execution has historically produced results with significant background banding that in many cases obscures the visualization of the target antigen. It has long been recognized that superior results can be achieved using primary anti-antigen/label conjugates in a western blot assays
Contents: 1 x 1 Kit of InstantBlot


InstantBlot - 1 Kit.jpg
 
InstantBlot technology lets you modify your primary antibody directly with HRP. No purification required. No more secondary antibodies needed resulting in reduced backgrounds. Each Instantblot kits converts up to 100 ¥ìg of primary antibody to a primary antibody-polyHRP conjugate in near quantitative yield, requiring only pipettes and a microcentrifuge. The whole process requires less than 30 minutes hands-on-time. With InstantBlot you obtain better results faster.

The final conjugate allows development of up to 40 western blots, which is dependent on the inherent affinity of the primary antibody.

Features

  • Minimal background
  • Faster procedures
  • Elimination of cross-species contamination
  • Multiplexing possibilitie
 

Actin detection in Mouse LLC lysate:

Mouse LLC lysate (10 ¥ìg to 0.625 ¥ìg) was transferred to nitrocellulose membrane and blocked with 4% BSA for 1 hour at room temperature
 


©¬-actin (43 kDa)





 
 
 
Lanes 1-5: Directdetection with InstantBlot
Blocked membranes were probed with mouse-actin-antibody-HRP conjugate (0.25 ¥ìg/mL) for 1 hour at room temperature. The target protein was visualised through ECL development.
Lanes6-9:Traditional detection with secondary antibodies
Blocked membranes were probed with mouse-actin-antibody-HRP conjugate (0.25 ¥ìg/mL) for 1 hour at room temperature. The blot was subsequently probed with goat antimouse-HRP conjugate (0.1 ¥ìg/mL) for 1 hour at room temperature; The target protein was visualised through ECL development.

Tubulin Detection in Mouse Splenocyte lysate:

InstantBlot                                                  Traditional
InstantBlot:
  1. Load sample 5 µg (Lane 1) to 2.5 µg (Lane 2).
  2. Transfer to nitrocellulosemembrane, 3% milk block, 1 hour.
  3. Incubate with 0.2 µg/mL ¥á-Tubulin antibody-HRP conjugate, 1 hour.
  4. ECL development
Traditional:
  1. Load sample 5 µg (Lane 1) to 2.5 µg (Lane 2).
  2. Transfer to nitrocellulosemembrane, 3% milk block, 1 hour.
  3. Incubate with 0.2 µg/mL ¥á-Tubulin antibody-HRP conjugate, 1 hour.
  4. Incubate with 0.15 µg/mL goat anti-mouse-HRP conjugate, 1 hour.
  5. ECL development

    Kit Components

    Components Quantity
    Antibody Activator 1 x 100 µg
    Activated-HRP 1 x 50 µL
    Modification Buffer 5 mL
    Spin Columns 3
    Collection Tubes 6
    DMF 0.2 mL
    Antibody Working Buffer 120 mL
    20X Wash Buffer 75 mL

     
     
    »ç¿ë Èıâ
    »ç¿ëÈıⰡ ¾ø½À´Ï´Ù.
    »ç¿ëÈıâ ÀÛ¼º
    gototop