The ultimate in Western blot sensitivity and speed.
Thermo Scientific Pierce Fast Western Blot Kit, SuperSignal West Femto Substrate contains optimized reagents and protocol for mid-to-low femtogram-level detection and typical Western blot development less than 1 hour after gel transfer.
The Fast Western Blot Kit protocol is a quick and efficient method for developing Western blots without the need to purchase expensive instrumentation and consumables required by other rapid Western blotting systems. These complete Pierce Fast Western Blot Kits require minimal hands-on time and yield results comparable to classic Western blotting with SuperSignal West Femto Chemiluminescent Substrate. The kits include SuperSignal West Femto Substrate, the most sensitive enhanced chemiluminescent substrate available for detecting horseradish peroxidase (HRP). This substrate’s sensitivity, intensity and duration of the signal allow for easy documentation using photographic or other imaging methods. The developed blots can be repeatedly exposed and stripped of the primary antibody and reprobed. The complete kit contains all the reagents necessary to block and probe a Western blot with either a mouse or rabbit primary antibody.
Highlights:
Fast – femtogram levels of detection in about 1 hour
Convenient – no expensive hardware or vacuum required; no clogging issues
Simple – optimized protocol makes Western blot analysis easier than ever
Easy – complete kits contain all components needed to block, probe and develop a blot with your mouse or rabbit primary antibody
Product Details:
Classic vs. fast Western protocol using Thermo Scientific SuperSignal West Femto Substrate. Cell lysate was diluted by 50% and separated by electrophoresis. Proteins were transferred to nitrocellulose and PVDF membranes using the Pierce Fast Semi-Dry Blotter for 10 minutes at 25V. Membranes were developed using either the classic Western blotting protocol or the fast Western protocol. Each blot was incubated in SuperSignal West Femto Substrate Working Solution for 5 minutes and exposed to film. Ezrin detection: HeLa cell lysates were probed with mouse anti-ezrin antibody (1µg/mL, Lab Vision) and exposed to film for 1 minute. Akt1 detection: 293T cell lysates were probed with mouse anit-Akt1 antibody (1/2000 dilution, Cell Signaling Technology) and exposed to film for 30 seconds.
Comparision of other Western blotting protocols to the Thermo Scientific Pierce Fast Western System. This table provides a comparison of the steps and times required to develop Western blot, after the transfer, using the Pierce Fast Western reagent-based system and instrument-based system from Millipore Corporation.
Protocol Steps
Classical protocol
Pierce Fast Western Blot Kit
SNAP i.d.*
Blocking/Pretreatment
60 min
- -
- -
Incubation with Primary
60 min
30 min
10 min
Wash
45 min
- -
1 min
Incubation with Secondary
60 min
10 min
10 min
Wash
45 min
15 min
1 min
Total Time
270 min
55 min
22 min
Key limitations
• Preparation of fresh non-fat dry milk blocking buffer each time
• For use with mouse or rabbit primary antibodies
• Only 2 full-sized blots per cycle. Strips limited by the format of the blot holders available
• Reliable vacuum source required
The Thermo Scientific Pierce Fast Western Blot Kit, SuperSignal West Femto Substrate is a cost-effective means to obtain Western blot results quickly. This table compares the cost of reagents and required consumables used in a classical Western blot protocol, the Pierce Fast Western Blot Kit reagent-based system and Millipore SNAP i.d. system.
Western blot reagents
Classical protocol
Pierce Fast Western System
SNAP i.d.
blot holder for instrument
- -
- -
$7.33
blotting membrane (NC)
$2.48
$2.48
$2.48
blocking buffer
$0.02
- -
$1.32
wash buffer (TBST)
$1.48
- -
$1.48
secondary antibody conjugate
$0.15
- -
$0.15
10ml detection substrate
(SuperSignal West Femto)
$26.65
- -
$26.65
film (5x7)
$1.05
$1.05
$1.05
kit's price per blot
- -
$33.60
- -
Total price per blot
$31.83
$38.93
$40.46
*Prices based on 2010 Thermo Scientific Pierce Western Blot Reagents and using SuperSignal West Femto Chemiluminescent Substrate as the detection reagent for comparable levels of sensitivity.
Fast Western Blot Kits, SuperSignal West Femto, Mouse
Formulation: Multi-component kit Sufficient For: 2 mini blots probed with mouse primary antibodies Kit Contents: Fast Western Antibody Diluent, 40mL Fast Western 10X Wash Buffer, 20mL Fast Western Mouse Optimized HRP Reagent, Femto, 2mL SuperSignal West Femto Luminol/Enhancer Solution, 10mL SuperSignal West Stable Peroxide Solution, 10mL
80mL kit
$106.00
35080
Fast Western Blot Kits, SuperSignal West Femto, Mouse
Formulation: Multi-component kit Sufficient For: 10 mini blots probed with mouse primary antibodies Kit Contents: Fast Western Antibody Diluent, 200mL Fast Western 10X Wash Buffer, 100mL Fast Western Mouse Optimized HRP Reagent, Femto, 10mL SuperSignal West Femto Luminol/Enhancer Solution, 50mL SuperSignal West Stable Peroxide Solution, 50mL
400mL kit
$384.00
35086
Fast Western Blot Kits, SuperSignal West Femto, Rabbit
Formulation: Multi-component kit Sufficient For: 2 mini blots probed with rabbit primary antibodies Kit Contents: Fast Western Antibody Diluent, 40mL Fast Western 10X Wash Buffer, 20mL Fast Western Rabbit Optimized HRP Reagent, Femto, 2mL SuperSignal West Femto Luminol/Enhancer Solution, 10mL SuperSignal West Stable Peroxide Solution, 10mL
80mL kit
$106.00
35081
Fast Western Blot Kits, SuperSignal West Femto, Rabbit
Formulation: Multi-component kit Sufficient For: 10 mini blots probed with rabbit primary antibodies Kit Contents: Fast Western Antibody Diluent, 200mL Fast Western 10X Wash Buffer, 100mL Fast Western Rabbit Optimized HRP Reagent, Femto, 10mL SuperSignal West Femto Luminol/Enhancer Solution, 50mL SuperSignal West Stable Peroxide Solution, 50mL
