Navigation Links
Application-Specific PVDF Membranes: Immun-Blot PVDF Membrane for Western Blotting and Sequi-Blot PVDF Membrane for Protein Sequencing, Rev C

Eric Larson, Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules, CA 94547 USA


Introduction
PVDF membrane was originally introduced to protein use as an ideal medium for the harsh chemical environment of N-terminal, or Edman degradation, sequencing. Even though PVDF is very hydrophobic and requires a prewetting step in alcohol, its high protein binding capacity, target retention, and resistance to cracking have made it an appealing membrane for general laboratory techniques.

The two main applications for PVDF are N-terminal sequencing and immunoblotting, both of which benefit from the qualities of PVDF but rely on different features of the membrane. While sequencing work is concerned with retaining as much protein as possible, a western blot requires good signal retention with very low background. To provide the best membrane for each technique, Bio-Rad offers two grades of PVDF: Immun-Blot PVDF membrane for western blotting and Sequi-Blot PVDF membrane for protein sequencing.


Immun-Blot PVDF Membrane
The focus with a blotting membrane is on how well it delivers signal while resisting background and nonspecific binding. Immun-Blot PVDF membrane is ideal for chemiluminescent (Figure 1) and colorimetric (Figure 2) western blots because it very strongly retains target protein (140150 g protein/cm2 membrane) but resists background that can obscure highsensitivity detection. Immun-Blot PVDF membrane retains proteins in any transfer format: tank blotting, semi-dry blotting, and dot blotting all deliver excellent results. For proteins that are difficult to transfer, up to 0.1% SDS can be added to the transfer buffer without affecting the binding of the proteins to PVDF. The results are consistently clean, easy-to-read blots. The physical strength of Immun-Blot PVDF membrane means that it will not crack or tear during common handling, and will hold up under repeated stripping and reprobing applications.


Sequi-Blot PVDF Membrane
For protein sequencing applications, Sequi-Blot PVDF membrane is the best choice due to its extremely high protein binding capacity of 170200 g/cm2. This is the original Bio-Rad PVDF membrane, designed to withstand the conditions of N-terminal sequencing while providing the binding capacity to sequence even low-abundance samples. The table shows the increased protein recovery possible with Sequi-Blot PVDF membrane. Recovery of blotted proteins is typically in the range of 80100% of the initial sample load, resulting in higher initial coupling yields.

The binding efficiency of Sequi-Blot PVDF membrane is illustrated in Figure 3. This experiment showed that Sequi-Blot PVDF membrane is able to retain proteins that transfer through a competitors product.


Bibliography
Hulmes JD et al., Strategies for microcharacterization of proteins using direct chemistry on sequencer supports, in Techniques in Protein Chemistry (T Hugli, ed) Academic Press, San Diego, 716 (1989)

Matsudaira P, Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes, J Biol Chem 262, 10,035 10,038 (1987)

Speicher DW, Microsequencing with PVDF membranes: efficient electroblotting, direct protein adsorption and sequencer program modifications, in Techniques in Protein Chemistry (T Hugli, ed) Academic Press, San Diego, 2435 (1989)

Speicher DW et al., Electrotransfer and sequence enhancements improve low picomole sequence analysis using PVDF membranes without glass fiber filters, J Prot Chem 9, 254255 (1990)

Towbin H et al., Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications, Proc Natl Acad Sci USA 76, 43504354 (1979)

Yuen SW et al., Microanalysis of SDS-PAGE electroblotted proteins, Biotechniques 7, 7483 (1989)


back to top
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Direct On-Membrane MS/MS Analysis using the Chemical Printer (CHIP-1000) and AXIMA-QIT
2. Imaging Chemiluminescent Western Blots
3. Detection of Phosphorylated and Total ERK and p38 MAPK by Bio-Plex Assay, ELISA, and Western Blotting, Rev A
4. Application of the Bio-Plex Cell Lysis Kit in Western Blot Analysis of Phosphoproteins, Rev A
5. Multiplex protein detection with the ECL Plex fluorescent Western blotting system using the Ettan DIGE Imager
6. Detection and quantitation of low-abundant proteins with ECL Plex fluorescent Western blotting
7. DNA/RNA Blotting With the Trans-Blot SD Semi-Dry Electrophoretic Transfer Cell
8. A New C-Terminal GST Vector for Protein Production in S. pombe
9. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
10. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
11. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:5/25/2016)... ... May 25, 2016 , ... Founder ... double board-certified in surgery and surgery of the hand by the National Board ... stranger to going above and beyond in his pursuit of providing the most ...
(Date:5/25/2016)... ... May 25, 2016 , ... Scientists at the University of ... tried for mesothelioma may be hampering the research that could lead to one good ... here to read it now. , The team evaluated 98 mesothelioma patients ...
(Date:5/25/2016)... ... , ... Lady had been battling arthritis since the age of two and ... Lady’s owner Hannah sought the help of Dr Jeff Christiansen of Superior Veterinary ... ligament and help with the pain of Lady’s arthritis. Dr Christiansen suggested that in ...
(Date:5/24/2016)... , ... May 24, 2016 , ... ... diabetes, and traumatic injuries, will be accelerated by research at Worcester Polytechnic Institute ... engines of wound healing and tissue regeneration. , The novel method, developed by ...
Breaking Biology Technology:
(Date:4/28/2016)... April 28, 2016 First quarter 2016:   ... 966% compared with the first quarter of 2015 The ... 589.1 M (loss: 18.8) and the operating margin was 40% (-13) ... Cash flow from operations was SEK 249.9 M (21.2) ... guidance is unchanged, SEK 7,000-8,500 M. The operating margin ...
(Date:4/26/2016)... DUBLIN , April 27, 2016 ... of the  "Global Multi-modal Biometrics Market 2016-2020"  report ... ) , The analysts forecast ... a CAGR of 15.49% during the period 2016-2020.  ... a number of sectors such as the healthcare, ...
(Date:4/15/2016)... , April 15, 2016  A new partnership ... more accurate underwriting decisions in a fraction of ... competitively priced and high-value life insurance policies to ... With Force Diagnostics, rapid testing (A1C, ... data readings (blood pressure, weight, pulse, BMI, and ...
Breaking Biology News(10 mins):