Obtain your Western blot results in about an hour without buying an instrument.
The Thermo Scientific Pierce Fast Western Blot Kit, ECL Substrate contains optimized reagents that shorten the time required to perform a typical chemiluminescent Western blot from 4 hours down to approximately 55 minutes.
The kit provides all the reagents necessary to complete a Western blot being probed with a mouse or rabbit primary antibody. The protocol requires minimal hands-on time and yields results comparable to classic Western blotting with ECL. The included Pierce ECL Substrate produces a chemiluminescent signal, which is detected using photographic or other imaging methods. Blots can be repeatedly exposed to film to obtain optimal results or stripped of the primary antibody and immunodetection reagents and reprobed.
Highlights:
Fast – all the sensitivity of Pierce ECL substrate and saving 4 to 5 hours per blot
Convenient – no expensive hardware or vacuum required; no clogging issues
Simple – optimized protocol makes Western blot analysis easier than ever
Economical – cost as little as $8 per blot; no expensive consumables or extra equipment
Excellent stability – kit is stable for 1 year stored at 4°C
Easy – complete kit gives you all the components you need to block and probe and develop a blot with your mouse or rabbit primary antibody
Product Details:
The Pierce Fast Western Blot Kit is reagent-based system that provides optimized reagents for blocking, antibody dilution and detection of Western blots with Pierce ECL substrate. The protocol is a quick, efficient and economic way to obtain Western blot results without the hassle of buying an instrument and the consumables required by other popular rapid Western blotting systems. No vacuum pump is needed so there are never any clogged lines or membranes that can occur when using instrument-based systems. Because there is no apparatus, there is no need to buy disposable blotting trays that limit the number of blots processed at one time.
Comparable results obtained with classic Western blotting protocol and Thermo Scientific Pierce Fast Western Blot Kit, ECL Substrate. The detection sensitivity for various target proteins was compared using the Fast Western Blot Kit and the classical Western blot protocol, Panels A and B contained 10, 2, 0.4 and 0.08µg of protein from A549 cell lysates in lanes 1-4, respectively. Panels C and D contained 2, 1, 0.5, 0.25 and 0.125µg of 293T cell lysate in lanes 1-5 of Panel C and 25, 12.5, 6.25, 3.1 and 0.39µg in lanes 1-5 of Panel D. Protein detection was achieved as follows: Panel A. Mouse anti-Cyclin D1 antibody (BD Pharmingen) on PVDF. Panel B. Rabbit anti-Beta Catenin antibody (LabVision) on PVDF. Panel C. Mouse anti-GAPDH antibody (Millipore) on nitrocellulose. Panel D. Rabbit anti-Cdk2 antibody (Upstate) on PVDF.
Researcher generated data using the Fast Western Blot Kit, ECL Substrate
Multiplex Experiment with Thermo Scientific Pierce Fast Western Blot Kit, ECL Substrate. C57BL/6 mice bone marrow cells were stimulated with M-CSF (50ng/ml) and RANKL (50ng/ml) to induce differentiation into osteoclasts. Total cell lysates on PVDF were probed with 0.2µg/m rabbit anti-hsp90 and mouse anti-cathepsin K (Santa Cruz Biotechnology). Data generated by Lai-Chu Wu, Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, Ohio.
Fast detection of His-tagged SAM68 protein purified using a Ni column from E. coli lysate.Lane 1. Lysate only, Lane 2. Column flow-through, Lanes 3-4. Wash and Lanes 5-8. Elution fractions. Fractions on PVDF were probed with rabbit anti-His-tag primary antibody (Santa Cruz Biotechnology). Data generated by Brenda Suh-Lailam and Joan Hevel, Department of Chemistry and Biochemistry, Utah State University, Logan, Utah.
Comparision of common Western blotting protocols to the Thermo Scientific Pierce Fast Western System. This table provides a side by side comparison of the steps and times required to complete a Western blot (post transfer and before development with substrate) using common classical methods, the Pierce Fast Western reagent-based system and instrument-based system from Millipore Corporation.
Protocol Steps
Classical protocol
Pierce Fast Western System
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 is a cost-effective means to obtain Western blot results quickly. This table provides a brief cost of reagents and required consumables used in a classical Western protocol, the Pierce Fast Western 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
detection substrate (ECL)
$1.43
- -
$1.43
film (5x7)
$1.01
$1.01
$1.01
kit's price per blot
- -
$7.92
- -
Total price per blot
$6.57
$11.41
$15.20
*Prices based on 2008 Thermo Scientific Pierce Western Blot Reagents.
Formulation: Multi-component kit Sufficient For: 25 mini blots probed with mouse or rabbit primary antibodies Kit Contents: Fast Western Antibody Diluent, 500mL Fast Western 10X Wash Buffer, 250mL Fast Western Optimized HRP Reagent, 25mL Pierce ECL Detection Reagent 1, 125mL Pierce ECL Detection Reagent 2, 125mL
1L kit
$216.00
35055
Pierce Fast Western Blot Kit, ECL Substrate
Formulation: Multi-component kit Sufficient For: 5 mini blots probed with mouse or rabbit primary antibodies Kit Contents: Fast Western Antibody Diluent, 100mL Fast Western 10X Wash Buffer, 50mL Fast Western Optimized HRP Reagent, 5mL Pierce ECL Detection Reagent 1, 25mL Pierce ECL Detection Reagent 2, 25mL
