Complete kit for multicolor fluorescent Western blotting

Detect two proteins on one blot using multicolor fluorescence, and gain more information from every experiment. With WesternBright MCF, you can assay a control alongside your protein of interest, or phosphorylated and unphosphorylated isoforms simultaneously. The WesternBright MCF fluorophores outperform Cy™ dyes, for high sensitivity and the ability to detect low pg amounts of protein. The kit contains all the materials needed to carry out 10 two-color fluorescent blots.

• Fluorescent detection of two proteins, with sensitivity 10x greater than Cy dyes
• AdvanBlock™ blocking solution for fast, protein-free blocking and greater signal-to-noise ratios
• High sensitivity – detect picograms of protein
• Fast results – entire protocol can be completed in 3.5 hours
• Compatible with imaging systems that detect Cy3 and Cy5 or similar dyes

WesternBright MCF fluorescent Western blotting kit allows the assay of two proteins at once, increasing the quality and quantity of information that can be gained from a single blot. Assay a loading control alongside a protein of interest. Assay phosphorylated and non-phosphorylated isoforms of a protein simultaneously. The WesternBright protocol saves time and money since there is no need to strip and re-probe a blot, no use of disposable film, and the blot can be imaged immediately, without drying.

The fluorescent dyes provided with WesternBright MCF outperform Cy dyes and allow detection of low pg amounts of protein.

 

Figure 1

Figure 1. WesternBright Conjugates provide a brighter signal than the ECL Plex Western Blotting system. Duplicate Western blots containing samples of AFP and CEA proteins were treated identically and probed with the same mixture of mouse anti-AFP and rabbit anti-CEA primary antibodies. One blot was then stained with WesternBright conjugates and the other with and identical concentration of Cy3 anti-mouse and Cy5 anti-rabbit antibodies, following the protocol recommended for ECL Plex. Under identical imaging conditions, WesternBright MCF provides a brighter signal, and 10x greater sensitivity.

 

Figure 2

Figure 2. Simultaneous detection of EGFR and phospho-EGFR with WesternBright MCF. The increase in phosphorylation of EGFR in response to EGF was detected. Lysates from A431 cells (lane 1) and from A431 cells treated with EGF (lane 2) were blotted and EGFR detected in the green channel (b) or phospho-EGFR detected in the red channel (c). The two channels are superimposed in (a). Lane 3: molecular weight markers.