Expedeon 2view™ double labeled secondary antibody
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- Protocols
- Specifications
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Innovative and unique detection method for Western blot
Expedeon’s 2view™ is an innovative and unique double labeled secondary antibody that enables extended detection within Western blotting. The product consists of a unique ternary complex developed using our world leading InnovaCoat® and Lightning-Link® conjugation technology. 2view™ secondary antibodies are labeled simultaneously with InnovaCoat® GOLD nanoparticles and horseradish peroxidase (HRP). The gold nanoparticles enable fast visible detection at nanogram level whereas the chemi- luminescent enzymatic detection enhances sensitivity down to picogram levels of protein.
Features & Benefits:
- Offers two modes of detection from a single antibody
- Provides highly sensitive chemi-luminescence analysis
- Easy to use with a single incubation
- Fast visible detection
- Dual level of sensitivity (nanogram and picogram level)
- High signal-to-noise ratio
Secondary antibodies available:
- Goat anti-Rabbit [Gold/HRP]
- Donkey anti-Goat [Gold/HRP]
- Goat anti-Mouse [Gold/HRP]
There are two levels of detection:
- Visible detection with InnovaCoat® GOLD
- Chemiluminescence via HRP & ECL Reagents (ECL Pico)
When performing a Western blot the amount of protein of interest is usually unknown; 2view™ solves this problem enabling two detections on the same blotted membrane: if the analyte is present down to nanogram level a red band will be visible; for lower amounts of analyte (picogram) the membrane can be developed using a chemiluminescent substrate (Expedeon’s LumiBlue ECL Pico Substrate).
1st level of detection – 40nm InnovaCoat® GOLD
Up to nanograms/single nanogram of analyte the detection relies on InnovaCoat® Gold nanoparticles and a red band will be visible by naked eye
2nd level of detection – HRP (ECL Pico)
Down to picograms of analyte the detection relies on the activity of HRP after the addition of ECL Pico (Expedeon). The signal can be recorded using a CCD camera or an X-Ray film
Comparison between 2view™ and traditional Western blot detection methods
Kit Contents:
- 10ml 2view™ anti-Rabbit / anti-Mouse/ anti-Goat [Gold/HRP]
Related Products:
- LumiBlue ECL solutions
- Electrophoresis Equipment
- RunBlue Protein Gels
- Buffers & Accessories for gel electrophoresis
- InstantBlue Protein Stain
- RunBlue Tris Glycine SDS Blot Buffer
- RunBlue Tris-Glycine Blot Buffer
- RunBlue Blot Membrane
For more information about this product please don’t hesitate to get in touch.
Innovative and unique detection method for Western blot
Expedeon’s 2view™ is an innovative and unique double labeled secondary antibody that enables extended detection within Western blotting. The product consists of a unique ternary complex developed using our world leading InnovaCoat® and Lightning-Link® conjugation technology. 2view™ secondary antibodies are labeled simultaneously with InnovaCoat® GOLD nanoparticles and horseradish peroxidase (HRP). The gold nanoparticles enable fast visible detection at nanogram level whereas the chemi- luminescent enzymatic detection enhances sensitivity down to picogram levels of protein.
Features & Benefits:
- Offers two modes of detection from a single antibody
- Provides highly sensitive chemi-luminescence analysis
- Easy to use with a single incubation
- Fast visible detection
- Dual level of sensitivity (nanogram and picogram level)
- High signal-to-noise ratio
Secondary antibodies available:
- Goat anti-Rabbit [Gold/HRP]
- Donkey anti-Goat [Gold/HRP]
- Goat anti-Mouse [Gold/HRP]
There are two levels of detection:
- Visible detection with InnovaCoat® GOLD
- Chemiluminescence via HRP & ECL Reagents (ECL Pico)
When performing a Western blot the amount of protein of interest is usually unknown; 2view™ solves this problem enabling two detections on the same blotted membrane: if the analyte is present down to nanogram level a red band will be visible; for lower amounts of analyte (picogram) the membrane can be developed using a chemiluminescent substrate (Expedeon’s LumiBlue ECL Pico Substrate).
1st level of detection – 40nm InnovaCoat® GOLD
Up to nanograms/single nanogram of analyte the detection relies on InnovaCoat® Gold nanoparticles and a red band will be visible by naked eye
2nd level of detection – HRP (ECL Pico)
Down to picograms of analyte the detection relies on the activity of HRP after the addition of ECL Pico (Expedeon). The signal can be recorded using a CCD camera or an X-Ray film
Comparison between 2view™ and traditional Western blot detection methods
Kit Contents:
- 10ml 2view™ anti-Rabbit / anti-Mouse/ anti-Goat [Gold/HRP]
Related Products:
- LumiBlue ECL solutions
- Electrophoresis Equipment
- RunBlue Protein Gels
- Buffers & Accessories for gel electrophoresis
- InstantBlue Protein Stain
- RunBlue Tris Glycine SDS Blot Buffer
- RunBlue Tris-Glycine Blot Buffer
- RunBlue Blot Membrane
For more information about this product please don’t hesitate to get in touch.
Innovative and unique detection method for Western blot
- Protocols Content
Sensitivity of gel and membrane staining methods
Figure 1. Transferrin (TF) was run on a 4-12% Bis-Tris gel (Expedeon, NBT41212), under non-reducing conditions for 40min at 180V in 1x MOPS (Expedeon, NXB75500). The gel was stained with InstantBlue™ (ISB1L) for one hour on the gel rocker (A). TF was blotted onto a nitrocellulose membrane (Expedeon, NXA19020), using 1x Tris-Glycine buffer (Expedeon, NXB86500) + 10% methanol. Membrane was reversibly stained with Ponceau S (B). After destaining, the membrane was blocked in 1x TBS, 0.05% Tween20 (Sigma, P1379-500ML), 5% BSA (VWR, 421501J) for 1 hour at RT on the gel rocker and then incubated O/N with anti-transferrin rabbit polyclonal diluted 1:5000 in 1x TBS, 0.05% Tween20 , 5% BSA on the gel rocker. Membrane was washed 3 times with 1x TBS, 0.05% Tween20 (5 minutes each wash on the gel rocker) and incubated for one hour with 2view™GAR [Gold/HRP] diluted 1:25 in blocking buffer (C).
Comparison of 2view™ GAR [Gold/HRP] and Goat anti-Rabbit HRP
Figure 2. Transferrin (TF) was run on a 4-12% Bis-Tris gel (Expedeon, NBT41212), under non-reducing conditions for 40’ at 180V in 1x MOPS (Expedeon, NXB75500). TF was blotted onto a nitrocellulose membranes (Expdeon, NXA19020), using 1x Tris-Glycine buffer (Expedeon, NXB86500) + 10% methanol. Membranes were blocked in 1x TBS, 0.05% Tween20 (Sigma, P1379-500ML), 5% BSA (VWR, 421501J) for 1 hour at RT on the gel rocker. Membranes were incubated O/N with anti-transferrin rabbit polyclonal diluted 1:5000 in 1x TBS, 0.05% Tween20 , 5% BSA on the gel rocker. Membranes were washed 3 times with 1x TBS, 0.05% Tween20 (5 minutes each wash on the gel rocker) and incubated for one hour with 2view™GAR [Gold/HRP] diluted 1:25 in blocking buffer and with Goat anti-Rabbit-HRP diluted 1:25.000 in blocking buffer on the gel rocker. Membranes were washed 3 times with 1x TBS, 0.05% Tween20 (5 minutes each wash on the gel rocker) and developed with ECL Pico (Expedeon, ECLP0250). Exposure time: 3s, 10s, 30s, 60s.
- Specifications Content
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