ONE-HOUR WesternTM Multiplex Fluorescent Kit (10 assays/kit)
ONE-HOUR WesternTM Multiplex Fluorescent Kit
Description:
GenScript ONE-HOUR WesternTM Multiplex Fluorescent Kit yields journal-quality Western results in about one hour. Using GenScript breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step Western process, which can take nearly five hours. Furthermore, this kit allows researchers to use multiple antibodies to detect multiple antigens on the same membrane at the same time. Transfer the proteins from gel to membrane and incubate it in the pretreat solution for five minutes. Then incubate in WB-M solution with primary antibodies and the corresponding fluorescent dye labeled secondary antibodies for 40 minutes. Then wash three times for ten minutes each. The membrane can then be scanned on a LI-COR Odyssey Infrared Imaging Systems.
The ONE-HOUR WesternTM Multiplex Fluorescent Kit is designed to produce high signal with low background for quick and clear Western analysis of multiple proteins. This kit can be used for any primary antibodies raised from these three species: rabbit, mouse, and goat. The corresponding fluorescent-dye labeled secondary antibodies are also needed.
Key Features
- Antibodies from all three major species (rabbit, mouse and goat) can be used.
- No cross-reaction.
- Reproducible results: The kit produces highly reproducible results.
Examples:
Multiplex Fluorescent Western blot detection of four proteins at the same time on the same membrane.
HeLa cell lysate was spiked with GST protein as shown in Figure 1. All the primary antibodies and secondary antibodies are listed in the following table.
| Antigens | Primary Antibodies | Amount | Secondary Antibodies | Amount |
|---|---|---|---|---|
| α-Tubulin |
Mouse Anti-α-Tubulin Monoclonal Antibody (Sigma, T6074) |
6 μg |
IRDye®680 Donkey Anti-Mouse (LI-COR, 926-32222) |
3 μg |
| β-Actin |
THETM Anti-β-actin Monoclonal Antibody (Mouse) (GenScript, A00702) |
6 μg |
IRDye®800CW Goat Anti-Mouse (LI-COR, 926-32210) |
3 μg |
| GAPDH |
Goat Anti-GAPDH Polyclonal Antibody (GenScript, A00191) |
4 μg |
IRDye®680 Donkey Anti-Goat (LI-COR, 926-32224) |
2 μg |
| GST |
Rabbit Anti-GST Polyclonal Antibody (GenScript, A00097) |
6 μg |
IRDye®800CW Goat Anti-Rabbit (LI-COR, 926-32211) |
3 μg |
Figure 1. Multiplex Fluorescent Western blots for the detection of α-Tubulin, β-Actin, GAPDH, and GST proteins using the ONE-HOUR WesternTM Multiplex Fluorescent Kit (L00398). A: 700 nm fluorescence image; B: 800 nm fluorescence image; C: The two fluorescence colors were imaged simultaneously in a single scan on a LI-COR Odyssey Infrared Imaging Systems. M is the Protein Marker for Fluorescent Western (GenScript, M00124).
Protein marker for Fluorescent Western
GenScript also provides protein marker for Fluorescent Western (M00124), the performance of the marker is demonstrated at below.
10 µl and 5 µl of reconstituted protein marker for Fluorescent Western blot were loaded in 12% Express SDS-PAGE Gel (GenScript, MG012W10).
Fig. A shows the results when IRDye 680-conjugated Goat Anti-Rabbit IgG (LI-COR) is used in the Western blot.
Fig. B shows the results when IRDye 800CW-conjugated Goat Anti-Mouse IgG (LI-COR) is used in the Western blot. Images were acquired using Odyssey Infrared Imaging system (LI-COR).
Technical Manual
Components
| Pretreat Solution A | 100 ml |
| Pretreat Solution B | 100 ml |
| WB-1 Solution | 2 ml |
| WB-M Solution | 100 ml |
| 10X Wash Solution | 125 ml |
| User Manual | 1 |
- SKU: L00398
- Size: kit
-
$170.50