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Immobilon® Membranes, Sandwiches and Blotting Filter Paper

Immobilon P membranes are the ideal PVDF transfer membranes for Western blotting

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Overview

Specifications

Ordering Information

Immobilon® Membranes, Sandwiches and Blotting Filter Paper Clear Sorting & Filtering
Catalog Numbericon Descriptionicon Filter Dimensionsicon Pack Sizeicon
IBFP0785CImmobilon Blotting Filter Paper, 7 x 8.4 cm sheet 7 cm x 8.4 cm 100 Show Pricing & Availability
IBFP0813CImmobilon Blotting Filter Paper, 8.5 x 13.5 cm sheet 8.5 cm x 13.5 cm 100 Show Pricing & Availability
IPFL00010Immobilon-FL PVDF, 0.45 µm, 26.5 cm x 3.75 m roll 26.5 cm x 3.75 m 1 Show Pricing & Availability
IPFL10100Immobilon-FL PVDF, 0.45 µm, 10 x 10 cm sheet 10 cm x 10 cm 10 Show Pricing & Availability
IPFL20200Immobilon-FL PVDF, 0.45 µm, 20 x 20 cm sheet 20 cm x 20 cm 10 Show Pricing & Availability
IPSN07852Blotting Sandwich Immobilon-P membrane interleaved with blotting paper 7 x 8.4 cm 7 cm x 8.4 cm 20 Show Pricing & Availability
IPSN08132Blotting Sandwich Immobilon-P membrane interleaved with blotting paper 8.5 x 13.5 cm 8.5 cm x 13.5 cm 20 Show Pricing & Availability
IPVH00010Immobilon-P Membrane, PVDF, 0.45 µm, 26.5 cm x 3.75 m roll 26.5 cm x 3.75 m 1 Show Pricing & Availability
IPVH07850Immobilon-P Membrane, PVDF, 0.45 µm, 7 x 8.4 cm sheet 7 cm x 8.4 cm 50 Show Pricing & Availability
IPVH08100Immobilon-P PVDF 0.45 µm 8 x 10 cm sheet 8 cm x 10 cm 10 Show Pricing & Availability
IPVH08130Immobilon-P PVDF 0.45 µm 8.5 x 13.5 cm sheet 8.5 cm x 13.5 cm 10 Show Pricing & Availability
IPVH09120Immobilon-P Membrane, PVDF, 0.45 µm, 9 x 12 cm sheet 9 cm x 12 cm 10 Show Pricing & Availability
IPVH10100Immobilon-P Membrane, PVDF, 0.45 µm, 10 x 10 cm sheet 10 cm x 10 cm 10 Show Pricing & Availability
IPVH15150Immobilon-P Membrane, PVDF, 0.45 µm, 15 x 15 cm sheet 15 cm x 15 cm 10 Show Pricing & Availability
IPVH20200Immobilon-P Membrane, PVDF, 0.45 µm, 20 x 20 cm sheet 20 cm x 20 cm 10 Show Pricing & Availability
IPVH304F0Immobilon-P Membrane, PVDF, 0.45 µm, 26 x 26 cm sheet 26 cm x 26 cm 10 Show Pricing & Availability
ISEQ00010Immobilon-PSQ Membrane, PVDF, 0.2 µm, 26.5 cm x 3.75 m roll 26.5 cm x 3.75 m 1 Show Pricing & Availability
ISEQ07850Immobilon-PSQ Membrane, PVDF, 0.2 µm, 7 x 8.4 cm sheet 7 cm x 8.4 cm 50 Show Pricing & Availability
ISEQ08100Immobilon-PSQ PVDF 0.2 µm 8 x 10 cm sheet 8 cm x 10 cm 10 Show Pricing & Availability
ISEQ08130Immobilon-PSQ PVDF 0.2 µm 8.5 x 13.5 cm sheet 8.5 cm x 13.5 cm 10 Show Pricing & Availability
ISEQ09120Immobilon-PSQ PVDF 0.2 µm 9 x 12 cm sheet 9 cm x 12 cm 10 Show Pricing & Availability
ISEQ10100Immobilon-PSQ Membrane, PVDF, 0.2 µm, 10 x 10 cm sheet 10 cm x 10 cm 10 Show Pricing & Availability
ISEQ15150Immobilon-PSQ Membrane, PVDF, 0.2 µm, 15 x 15 cm sheet 15 cm x 15 cm 10 Show Pricing & Availability
ISEQ20200Immobilon-PSQ Membrane, PVDF, 0.2 µm, 20 x 20 cm sheet 20 cm x 20 cm 10 Show Pricing & Availability
ISEQ26260Immobilon-PSQ Membrane, PVDF, 0.2 µm, 26 x 26 cm sheet 26 cm x 26 cm 10 Show Pricing & Availability

*Roll

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    Documentation

    Brochure

    Title
    Western Blotting Tools

    Technical Info

    Title
    Protein Blotting Handbook: 6th Edition
    Protein Dot Blotting using Immobilon-P

    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)
    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
    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

    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
    Characterization of retinoic acid receptor-deficient keratinocytes.
    Goyette Philippe; Chen Chang Feng; Wang Wei; Seguin Francois; Lohnes David(a)
    Journal of Biological Chemistry v 275 pg 16497-16505 June 2, 2000  2000

    Protection of renal inner medullary epithelial cells from apoptosis by hypertonic stress-induced p53 activation
    Dmitrieva Natalia(a); Kultz Dietmar; Michea Luis; Ferraris Joan; Burg Maurice ; Journal of Biological Chemistry v 275, pg 18243-18247 June 16, 2000
    Journal of Biological Chemistry v 275, pg 18243-18247 June 16, 2000  2000

    FAQ

    QuestionAnswer
    Should I prewet MultiScreen Immobilon-P plates before use? Yes. Use 15 ul of 70% ethanol or methanol to prewet the Immobilon P membrane. This membrane is hydrophobic and requires a prewet to allow liquid to pass through the membrane.
    What is the thickness of the Immobilon PSQ? Immobilon PSQ is 200 microns thick.
    What is the smallest size of a protein or peptide which binds to the Immobilon-PSQ? For the PSQ we do not suggest that they go lower than 2 kDa for the protein size. To help promote the transfer of the smaller proteins the methanol concentration of the transfer buffer can be increased to 20% (w/v) and the SDS lowered to 0.01%. The strength of the electric field can also be reduce by 50% to increase the proteins contact time with the membrane. Standard stains can be used such as Coomasie Blue and Ponceau. Keep in mind that some of the stain such as Coomasie Blue are not reversible.
    How many times can I strip and reprobe Immobilon-P? While 2-3 times is probably the limit, it is difficult to give an absolute number in regards to stripping and reprobing. This is because there are many factors to consider when it comes to protein binding to PVDF and primary antibody affinity to these proteins. Different proteins will have varying degrees of affinity to PVDF. This is based on factors discussed in our Protein Blotting Handbook (see TP001; Protein Binding section). One round of stripping may remove one protein and leave another intact. The same could occur when using different primary antibodies. Different antibodies will have varying affinities to different proteins. If carrying out several rounds of stripping and reprobing, one strategy might be to detect the least abundant proteins earlier leaving the higher abundant proteins later for detection.
    What is the difference between Immobilon–FL and Immobilon-P? Both membranes are made from PVDF. The difference in background fluorescence is due to proprietary modifications in the membrane manufacturing process.
    What fluorescence-based detection methods can be used with Immobilon-FL? Immobilon-FL can be used in Western blotting applications using either fluorescent dye-conjugated antibodies or chemi-fluorescence substrates. Western blots can be imaged in the visible or IR range. Single color detection or multiplexing for co-localization studies can be performed efficiently on Immobilon-FL Fluorescent proteins, e.g. green fluorescent protein, (GFP) or proteins tagged with GFP, blotted onto the membrane can be readily detected as well.
    What fluorescent dyes are recommended to use with Immobilon-FL? Immobilon-FL membrane exhibits very low background fluorescence over a broad range of wavelengths and can be used with all visible and infrared fluorescent dyes. Generally, dyes with larger Stokes shift (greater separation between absorption and emission wavelengths) are preferred in western blotting applications using fluorescence-based detection. NOTE: To prevent photo-bleaching, protect the membrane from light during secondary antibody incubations and washes until the membrane is ready to be scanned.
    How long does the fluorescent signal stay on the developed blot? The fluorescent signal from fluorescence-tagged antibodies will remain stable on the membrane for several months, or longer, when stored protected from light. The membranes may be stored dry or in PBS at 4ºC. The fluorescent signal from chemi-fluorescent substrates does not remain for long so blots using these substrates need to be scanned immediately.
    Do I need to dry Immobilon-FL blots before scanning? Drying the blot may enhance signal strength. NOTE: Do not wrap the blot in plastic/saran wrap while scanning. If you do need to wrap the membrane, Mylar should be used.
    Can Immobilon-FL be stripped and re-probed? Yes, Immobilon-FL can be stripped and re-probed like any other PVDF membrane, using standard detergent or acidic methods (refer to Millipore’s Protein Blotting Handbook). Keep the blot wet. If dried, rewet with methanol and then rehydrate before stripping the blot. Once the membrane has dried the stripping is ineffective. Alternatively, the blot can be photo-bleached to remove the fluorescence.

    User Guides

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