Transform your SpectraMax i3x or SpectraMax Paradigm System into a western blot reader
There are often a great many western blots along the path between imagination and understanding. Now you can read them using the advanced optical technology that’s already in your lab — inside your microplate reader. The ScanLater™ Western Blot Detection System includes the ScanLater Western Blot Detection Cartridge, ScanLater Western Blot Assay Kit, and image acquisition powered by SoftMax® Pro Software. In just minutes, you can install the ScanLater Western Blot Detection Cartridge in a SpectraMax® i3x or SpectraMax® Paradigm® System to add western blot detection capability, rather than buying a dedicated western blot detection system. See more, and do more, with your microplate reader.
- Overview
- Technology
- Data
- Resources
The ScanLater Western Blot Detection Cartridge is a user-installable option for the SpectraMax i3x and SpectraMax Paradigm Multi-Mode Platforms. Leveraging this novel technology, researchers can:
- Add western blot detection to their microplate reader within 2 minutes
- Eliminate time-dependent substrate addition steps
- Maintain femtogram to picogram protein sensitivity comparable to traditional western blot methods
- Sustain blot signal stability for up to 30 days
- Use a single software platform SoftMax Pro Software, to run both microplate and western blot detection assays
Features:
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- User-installable cartridge for the SpectraMax i3 and Paradigm Multi-Mode Platforms
- Time-resolved fluorescence detection
- Xenon flash lamp excitation source 340/80 nm excitation range
- 616/10 nm emission range
- Supports nitrocellulose and PVDF membranes
- Compatible with goat anti-rabbit, anti-mouse or streptavidin labeled antibodies
- Reads membranes fitting the ScanLater SpectraMax Membrane Holder
Figure 1.
ScanLater™ Western Blot Assay Workflow
Figure 1. The ScanLater System simplifies the traditional western blot protocol
by employing a secondary antibody labeled with a europium-chelate.
The blot is then read using the ScanLater Western Blot Cartridge on the
SpectraMax® i3 or Paradigm Multi-Mode Platforms.
No substrates are needed and the blot can be scanned immediately after washing.
Figure 2.
ScanLater Western Blot Assay Principle
Figure 2. Schematic of protein detection method shows the
secondary antibody is directly labeled with Europium.
Figure 3.
Europium-chelate excitation and emission spectra
Figure 4.
Schematic representation of emission lifetimes that minimize background fluorescence
Figure 4. Schematic representation of emission lifetimes of background fluorescence
and Eu-chelate showing principle of TRF measurement.
Signal is measured after appropriate time-delay to reduce auto-fluorescence.
Representative applications:
- Protein identification and quantitation
- Epitope mapping
- Amino acid composition and sequence analysis
- Spots imprinting
- Phosphoprotein endogenous or exogenous expression
- Protein resillience
- Structure domain analysis
- Protein expression
- Protein content in serum
- QC elimination of albumin and lgG in serum
- Protein expression the cell cycle
ScanLater Western Blot System sensitivity
and dynamic range
Figure 1. Image of Gluthathione S-transferase dilution series as scanned by the SpectraMax Paradigm Reader. Note: False-color scale is used to show large dynamic range. Integrated intensities from individual bands showing total dynamic range of greater than 3 logs.
Comparison of ScanLater Western Blot System to chemiluminescent western blot detection
Figure 2. Detection of ubiquitinated Rad-18 in HEK 293 cells on treatment with different concentration (0, 50, 100 ppm) of carcinogen MMS, an alkylating agent. Comparable result using ScanLater Western Blot System vs. chemiluminescence western blot (Stanford University).