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Immobilon Western Substrates are optimized for high sensitivity and low background over a broad dynamic range at a considerably lower cost than other chemiluminescent reagents.
The substrates' high signal intensity allow detection of high and low abundance proteins. Antibody dilutions can usually be significantly increased compared to other chemiluminescent substrates, which reduces antibody consumption and assay costs.
Immobilon Western Substrates excel in all Western or dot/slot/spot blotting applications. They are compatible with both PVDF and nitrocellulose membranes, as well as all commonly used buffers and blocking reagents.
|Emission Duration||>2 hours||
|Primary Detection Method||X-ray film or imaging equipment||X-ray film or imaging equipment|
|Typical Antibody Dilution*||
>9 months at time of receipt
Working solution: 10 days at 2-8°C
|>9 months at time of receipt|
|Recommended Initial Exposure Time for Film||30 seconds||30 seconds|
|Membrane Compatibility||PVDF or nitrocellulose||PVDF or nitrocellulose|
Higher Sensitivity than Other AP Substrates Detection of transferrin using three AP substrates on Immobilon-P Membrane (Millipore).
Hydroxide ion (OH-) generated by conjugated horseradish peroxidase (HRP) in aprotic media result in transitions of luminol to 3'-aminophthalate, which induce emission of 425-510 nm light. To detect the induced emission of 425-510 nm light from the Spray & Glow™ reagent; use either a darkroom and standard X-ray film or a digital imaging technology.
Before ReBot Plus: Western blot probed with rabbit antibodies to peptide from angiotensin II type 1 receptor, 20 second exposure.
After ReBot Plus: Western blot stripped with ReBlot Plus Mild stripping solution.
Reprobe: Western blot reprobed with rabbit antibodies to peptide from glutamate receptor delta 1/2, 20 second exposure.
This kit contains two specially formulated antibody stripping solutions (Mild and Strong) that quickly and effectively remove antibodies and their corresponding chemiluminescent signal from membrane blots, without destroying the blotted protein sample.
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