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:2/9/2016)... ... February 08, 2016 , ... Date and ... Blumberg Institute at the Pennsylvania Biotechnology Center of Bucks County, 3805 Old Easton ... Medical College (TCMC) will hold an open house for participants to learn about ...
(Date:2/9/2016)... ... ... The American Academy of Thermology (AAT) has announced that for ... Certification Qualification Course for Technicians via a two part webinar on July 30 and ... review of hardware, software, and camera setup/operations, aligns with the in-person member qualification course ...
(Date:2/9/2016)... 9, 2016 This market research report on ... future prospects of the market in terms of revenue ... engaged in the manufacture of microbiology culture media and ... a market snapshot providing the overall information of various ... This section also provides the overall information and data ...
(Date:2/9/2016)... , Feb. 9, 2016 ... Inhibitors-Pipeline Insights, 2016", report provides in depth ... development activities around the Protein-Tyrosine Phosphatase 1B ... product profiles in various stages of development ... II, Phase III and Preregistration. Report covers ...
Breaking Biology Technology:
(Date:2/2/2016)... , Feb. 2, 2016 Technology Enhancements Accelerate ... an analysis of the digital and computed radiography markets ... , and Indonesia (TIM). ... and market size, as well as regional market drivers ... and discusses market penetration and market attractiveness, both for ...
(Date:2/1/2016)...  Wocket® smart wallet ( www.wocketwallet.com ) announces the launch of a ... . Las Vegas , where Joey appeared at ... Las Vegas , where Joey appeared at the Wocket booth ... ad was filmed at the Consumer Electronics Show (CES2016) in ... meet and greet fans. --> --> ...
(Date:1/22/2016)... 22, 2016 ... of the  "Global Behavioral Biometric Market ... --> http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ) has announced ... Biometric Market 2016-2020"  report to their ... Markets ( http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ) has announced the ...
Breaking Biology News(10 mins):