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Immobilon® Membranes, Sandwiches and Blotting Filter Paper

Immobilon® PVDF membranes are the ideal transfer membranes for protein blotting applications.

Related Resources

Overview

Specifications

Ordering Information

Immobilon® Membranes, Sandwiches and Blotting Filter PaperClear Sorting & Filtering Show Filter
Catalog NumberDescriptionFilter DimensionsPack Size
IPVH07850Immobilon®-P PVDF Membrane 7 cm x 8.4 cm 50 Show Pricing & Availability
IPVH08100Immobilon®-P PVDF Membrane 8 cm x 10 cm 10 Show Pricing & Availability
IPVH08130Immobilon®-P PVDF Membrane 8.5 cm x 13.5 cm 10 Show Pricing & Availability
IPVH09120Immobilon®-P PVDF Membrane 9 cm x 12 cm 10 Show Pricing & Availability
IPVH10100Immobilon®-P PVDF Membrane 10 cm x 10 cm 10 Show Pricing & Availability
IPVH15150Immobilon®-P PVDF Membrane 15 cm x 15 cm 10 Show Pricing & Availability
IPVH20200Immobilon®-P PVDF Membrane 20 cm x 20 cm 10 Show Pricing & Availability
IPVH304F0Immobilon®-P PVDF Membrane 26 cm x 26 cm 10 Show Pricing & Availability
IPVH00010Immobilon®-P PVDF Membrane 26.5 cm x 3.75 m 1 Show Pricing & Availability
ISEQ07850Immobilon®-PSQ PVDF Membrane 7 cm x 8.4 cm 50 Show Pricing & Availability
ISEQ08100Immobilon®-PSQ PVDF Membrane 8 cm x 10 cm 10 Show Pricing & Availability
ISEQ08130Immobilon®-PSQ PVDF Membrane 8.5 cm x 13.5 cm 10 Show Pricing & Availability
ISEQ09120Immobilon®-PSQ PVDF Membrane 9 cm x 12 cm 10 Show Pricing & Availability
ISEQ10100Immobilon®-PSQ PVDF Membrane 10 cm x 10 cm 10 Show Pricing & Availability
ISEQ15150Immobilon®-PSQ PVDF Membrane 15 cm x 15 cm 10 Show Pricing & Availability
ISEQ20200Immobilon®-PSQ PVDF Membrane 20 cm x 20 cm 10 Show Pricing & Availability
ISEQ26260Immobilon®-PSQ PVDF Membrane 26 cm x 26 cm 10 Show Pricing & Availability
ISEQ00010Immobilon®-PSQ PVDF Membrane 26.5 cm x 3.75 m 1 Show Pricing & Availability
IPFL10100Immobilon®-FL PVDF Membrane 10 cm x 10 cm 10 Show Pricing & Availability
IPFL20200Immobilon®-FL PVDF Membrane 20 cm x 20 cm 10 Show Pricing & Availability
IPFL00010Immobilon®-FL PVDF Membrane 26.5 cm x 3.75 m 1 Show Pricing & Availability
IPSN07852Immobilon®-P Blotting Sandwich 7 cm x 8.4 cm 20 Show Pricing & Availability
IPSN08132Immobilon®-P Blotting Sandwich 8.5 cm x 13.5 cm 20 Show Pricing & Availability
IBFP0785CImmobilon® Blotting Filter Paper 7 cm x 8.4 cm 100 Show Pricing & Availability
IBFP0813CImmobilon® Blotting Filter Paper 8.5 cm x 13.5 cm 100 Show Pricing & Availability
IPFL00005Immobilon®-FL PVDF Membrane 26.5 cm x 1.875 m 1 Show Pricing & Availability
IPVH00005Immobilon®-P PVDF Membrane 26.5 cm x 1.875 m 1 Show Pricing & Availability
ISEQ00005Immobilon®-PSQ PVDF Membrane 26.5 cm x 1.875 m 1 Show Pricing & Availability
IEVH00005Immobilon®-E PVDF Membrane 26.5 cm x 1.875 m 1 Show Pricing & Availability
IEVH07804Immobilon®-E PVDF Membrane 7 cm x 8.4 cm 4 Show Pricing & Availability
IEVH07850Immobilon®-E PVDF Membrane 7 cm x 8.4 cm 50 Show Pricing & Availability
IEVH08100Immobilon®-E PVDF Membrane 8 cm x 10 cm 10 Show Pricing & Availability
IEVH09120Immobilon®-E PVDF Membrane 9 cm x 12 cm 10 Show Pricing & Availability
IEVH10100Immobilon®-E PVDF Membrane 10 cm x 10 cm 10 Show Pricing & Availability
IESN07852Immobilon®-E Blotting Sandwich 7.0 cm x 8.4 cm 20 Show Pricing & Availability
IESN08132Immobilon®-E Blotting Sandwich 8.5 cm x 13.5 cm 20 Show Pricing & Availability
IMDISPImmobilon® NOW Dispenser 1 Show Pricing & Availability
IPVH85RImmobilon®-P Membrane, PVDF, 0.45 µm, 8.5 cm x 10 m roll 8.5 cm x 10 m 1 Show Pricing & Availability
ISEQ85RImmobilon®-PSQ Membrane, PVDF, 0.2 µm, 8.5 cm x 10 m roll 26.5 cm x 3.75 m 1 Show Pricing & Availability
IEVH85RImmobilon®-E PVDF Membrane 8.5 cm x 10 m 1 Show Pricing & Availability
IPFL07810Immobilon®-FL PVDF Membrane 7 cm x 8.4 cm 10 Show Pricing & Availability

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Documentation

Brochure

Title
Western Blotting Tools

Technical Info

Title
Protein Blotting Handbook: 6th Edition

Data Sheet

Title
Immobilon Transfer Membranes: For superior protein and nucleic acid blots

Posters

Title
History of Western Blotting
Low Background Membrane for Fluorescent Protein Detection in Western Blotting

References

Reference overviewApplication
Mobile Phase Preparation for UHPLC: Membrane Filtration Method Affects System Performance and Leaching of Extractable Impurities
Subodh Kulkarn(1), Jesmi George(2) and Vivek Joshi(2) (1) Millipore India Pvt. Ltd., Bioscience Division, 50A, 2nd Phase, Ring Road, Peenya, Bangalore, India 560058 (2) Millipore Corp., Bioscience Division, 17 Cherry Hill Drive, Danvers, MA 01923
LCGC  2010

Show Abstract Full Text Article
Quantitation of Protein on gels and blots by infrared fluorescence of Coomassie blue and fast green
Luo S., Wehr N.B., Levine R.L.
Analytical Biochemistry:350 (2006):233-238  2006

Immunoblotting (Western)
Role of the Small Heat Shock Proteins in Regulating Vascular Smooth Muscle Tone
McLemore E.C., Tessier D.J., Thresher J., Komalavilas P., Brophy C.M
J. Am. Coll. Surg. 2005, Vol 201 (1):30-36  2005

Pre-B-cell colony-enhancing factor is a secreted cytokine-like protein from the human amniotic epithelium.
Ognjanovic S, Ku TL, Bryant-Greenwood GD.
Am J Obstet Gynecol. 2005 Jul;193(1):273-82  2005

Western Blotting
A high-affinity reversible protein stain for Western blots
Antharavally B.S., Carter, B., Bell, P.A., Mallia K.
Analytical Biochemistry 2004,Vol 329:276-280  2004

Biochemical analysis of GABA receptor subunits alpha 1, alpha 5, beta1 beta2 in the hippocampus of patients with Alzheimer's disease neurophathology
Rissman, R.A., Mishizen-Eberz A.J. N.,Wolfe, C.B.B., DeBlas A.L., Miralles C.P., Ikonomovic M.D., armstrong D.M.
Neuroscience 120 (2003) 295-705  2003

Western Blotting
Proteomics reveals protein profile changes in doxorubicin treated MCF7 human breast cancer cells
Cheng S.T., Pan T, L., Tsai Y.Ch, Huang C. M.
Cancer letters 2002. vol 181:95-107  2002

Towards proteome-wide production of monoclonal antibody by phage display.
Bin Liu, Lan Huang, Carina Sihlbom, Al Burlingame and James D. Marks.
J Mol Biol. 2002 Feb 1;315(5):1063-73  2002

Mass Spectrometry Sample Prep
The clinical usefulness of the measurement of cytokines.
Bienvenu J et al.,Clin Chem Lab Med. 2000 Apr;38(4):267-85. Review.
Clin Chem Lab Med. 2000 Apr;38(4):267-85. Review.  2000

ELISPOT Assays
Amino Acid Analysis: A comparative Study of stains on PVDF membranes
,Journal of Biomolecular Techniques, Vol II, Issue 1, March 2000.
Journal of Biomolecular Techniques, Vol II, Issue 1, March 2000.  2000

FAQ

QuestionAnswer
I transferred my protein to Immobilon-P and I can't find it. I've stained the gel and the membrane and nothing is there. What happened?If the pI of your proteins are greater than the pH of the transfer buffer, the proteins will travel in the opposite direction. The protein probably transferred into the running buffer. If you suspect that your protein has a high pI, try CAPS buffer and/or put membrane on both sides of the gel.
How can I elute protein off Immobilon-P?Several methods have been used depending on the nature of the study. Proteins can be eluted from the membrane by incubation at room temp. in 50 mMTris-HCL, pH 9, containing 2% SDS and 1% Triton X-100, followed by dialysis to remove most of the detergents. If the eluted protein is to be purified by HPLC, hydrogenated Triton-X-100 should be used as it will not contribute any UV absorbence to the chromatogram. If detergents in the protein solution are a problem, a 40% acetonitrile solution in 0.1 M ammonium acetate, pH 8.9 for 3 hours at 37oC can be used to elute the proteins. Lyophilization of the extract will remove the volatile solvents. If the eluted protein is to be purified by HPLC, hydrogenated Triton-X-100 should be used as it will not contribute any UV absorbence to the chromatogram.
If I do a dot blot with Immobilon-P should I prewet the membrane?A dot blot can be done in two ways, via vacuum filtration with a dot blot apparatus or through intrusion with a tuberculin syringe. With the first method, the membrane must first be rendered hydrophilic using methanol (this allows the protein to adsorb onto the membrane) followed by a quick soak in water prior to assembly in the apparatus. When using the second method it isn't necessary to prewet with methanol. The user simply holds the syringe directly against the membrane and lets the pressure exerted from the syringe intrude the solution ( and protein) onto the membrane.
What is the sensitivity of the transillumination method with Immobilon-P and how does it work?The sensitivity of transillumination is equivalent to Coomassie Blue staining. After drying the membrane, the PVDF is re-wet in 20% methanol and viewed in white light, or placed in a tray containing 20% methanol on a white light transillumination box.

This method takes advantage of the change in refractive index of the rehydrated protein band to that of the water/methanol solution. The naturally hydrophobic Immobilon P membrane will not re-wet and, therefore can be visualized by the difference of refractive indices. The protein pattern appears translucent against the white membrane.
How can I strip Immobilon-P?Processing a single blot with multiple antibodies requires the removal of the first antibody prior to the addition of a second antibody preparation. The stripping method should effectively disrupt the antigen-binding capacity of the antibody and solubilize it into the surrounding buffer. For most immunodetection procedures, it is also important to prevent spurious binding of the secondary antibody during stripping so that the enzyme conjugate will not contribute to background in subsequent rounds of detection. Finally, the stripping method should not cause a loss of antigens from the membrane. Two methods are provided for stripping an immunoblot. http://www.millipore.com/publications.nsf/docs/tp001en00protocols.
Can I use Coomasie Blue G-250 to stain my protein blots instead of Coomasie Blue R-250 (Brilliant Blue) which is recommended in your literature?Coomasie Blue G-250 will work, however, be aware that it will not have the same intensity and contrast as the R-250 will. The difference is caused by the way the stain interacts with the protein. G-250 doesn't give the same level of binding as the R-250. Bands may look light with the G-250, but keep in mind that the protein could still be there at a higher concentration than the stain would indicate.
Can Ponceau-S be used to stain proteins transfered to Immobilon-P?Ponceau-S can be used with Immobilon-P and is reversible. The stain produces pinkish bands on a light background. The stain can be removed by washing the blot with 0.1N NaOH.
What are the important factors to consider when choosing a stain for use with Immobilon-P?The most important factor in choosing a stain is how the proteins on the blot are to be analyzed. For example, if immunodetection analysis will follow, non-reversible stains are not recommended. Non-reversible stains generally exhibit the best sensitivity but can interfere with or prevent further analysis of the proteins. Although less sensitive, reversible stains allow assessment fo the blot and then can be washed from the membrane. However, there is a risk that some proteins may undergo chemical modification during the process or be washed from the membrane.
My pre-stained protein markers disappear when I transfer my Western blot. What's happening?Some of the dyes used in pre-stained markers are very hydrophilic and the dyed protein may travel faster and penetrate more deeply into the membrane than the undyed protein. You can still check for the presence of your protein by transillumination of the membrane (after blot is dry, soak in 20% methanol for 5 minutes, and view translucent protein bands on a light box).
I want to strip my immunoblot and probe with another antibody. The rapid immunodetection protocol states that the membrane should be dry, but the stripping protocol states that the membrane should not be allowed to dry. What should I do?You dry the blot after transfer, and perform your rapid immunodetection protocol. Then, if you are planning to do rounds of immunodetection with other antibodies, you must keep the the blot wet after immunodetection is complete and continue with stripping off the first antibody. After that, you can proceed to the blocking step of the next round of immunodetection with the other antibody. Do not let the blot dry between rounds of immunodetection, otherwise, any residual antibodies from the first round will bind permanently to the membrane.

User Guides

Title
Immobilon -P Blotting Sandwiches
Immobilon®-E Transfer Membrane User Guide
Immobilon®-P Transfer Membrane User Guide
Re-Blot Plus Western Blot Recycling Kit