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RFP Western Blot Kit: for RFP Chemiluminescent Western Blotting

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RFP Western Blot: Known amounts of recombinant RFP and GFP protein (p/n 000-001-215) were spiked into a HeLa cell-derived lysates (p/n W09-000-364) and separated by SDS-PAGE using a 4-20% gradient gel. Proteins were transferred onto a nitrocellulose membrane for 1 h at 100 mV. The membrane was blocked with TTBS (p/n MB-013) supplemented with 1% BSA (p/n BSA-50) for 1 h at 4°C prior to probing the blot with the anti-GFP monoclonal antibody (p/n 600-301-215) diluted 1:1,000 for 40 min. Detection of the primary antibody by the HRP-conjugated anti-Mouse IgG (p/n 610-4302) was performed at a dilution of 1:20,000 for 1h at 4°C.  FemtoMax Super Sensitive Chemiluminescent Luminol Substrate was used for signal detection (see below).
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RFP Western Blot Kit: for RFP Chemiluminescent Western Blotting Properties

RFP Western Blot Kit: for RFP Chemiluminescent Western Blotting - KCA379
Known Cross Reactivity
wt RFP from Discosoma spp, rRFP, mCherry, tdTomato, mBanana, mOrange, mPlum, mOrange and mStrawberry
Western Blot : YES
Other Dilution: User Optimized
Product Type
Kit Type
Chemiluminescent Western Blot Kit
Physical State
Shipping Condition
Wet Ice
Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity!
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RFP Western Blot Kit: for RFP Chemiluminescent Western Blotting Description

Red Fluorescent Protein (RFP) Western Bloting kit allows for the detection of RFP-tagged recombinant proteins present in cell lysates provided by the user. After protein separation by SDS-PAGE and transfer, the membrane is probed with Rockland's optimized Anti-RFP antibody. Detection of the membrane bound antibody-antigen complex is achieved by the addition of a secondary antibody conjugated to the enzyme horseradish peroxidase. The enzyme reacts with a specialized formulation of luminol, an extremely sensitive, non-radioactive substrate that emits light and allows visualization using X-ray film or other imaging methods, including highly sensitive CCD cameras and imaging systems.
Western Blotting Kit, Chemiluminescent Kit, Peroxidase Kit, Immunoblotting Kit, RFP, rRFP, mCherry, tdTomato, mBanana, mOrange, mPlum, mOrange and mStrawberry
Storage Condition
See kit insert for complete instructions.
Application Note
Use Rockland Immunochemicals’ Anti-RFP Chemiluminescent Kit for Western Blotting for detection of RFP-tagged recombinant proteins by western blot. This kit is useful for both western blotting and dot blotting methods. Please read the entire product insert prior to use.
Rockland Immunochemicals’ Chemiluminescent Western Blot Kit for RFP combines all of the necessary reagents with a rapid proven protocol and the extremely high signal detection of our luminol chemiluminescent substrate for the detection of recombinant proteins containing wild type Red Fluorescent Protein (DsRed or RFP) and its variants such as mBanana, mCherry, DsRed2, E2-Crimson, mOrange, mOrange2, mPlum, mRaspberry, mStrawberry and tdTomato. The Chemiluminescent Western Blot Kit design includes straightforward procedures and color-coding to allow for ease of use. This kit contains sufficient substrate for up to 30 mini blots at 7.5 x 8 cm2 (1,800 cm2) and is stable for at least 1 year when stored as indicated.
Disclaimer Note-General
This material is subject to proprietary rights and is sold under license from ROCKLAND IMMUNOCHEMICALS Inc. This product is licensed for sale only for 'research-use' only. There is no implied license hereunder for any commercial use. FemtoMax® is a trademark of ROCKLAND IMMUNOCHEMICALS Inc. This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Antibodies, A Laboratory Manuel. Ed Harlow and David Lane, eds. Cold Spring Harbor Press. 1988. Chapter 12 gives an excellent overview of Western Blotting techniques, including India Ink staining. Current protocols in Molecular Biology. J. Ausebel, et al, eds. John Wiley and Sons, New York. Gives a complete protocol of Western Blotting and Dot Blotting. Molecular Cloning: A Laboratory Manuel. 2nd Edition. J. Sambrook, E.F. Fritsch and T. Maniatis, eds. Cold Spring Harbor Press, 1989. Chapter 18 gives detailed protocols for both the production of cell lysates and electrophoresis and blotting of proteins. Antibodies, A Practical Approach. 2nd Edition. Catty, D., ed. IRL Press, Oxford, England. 1990. Volumes I and II represent a detailed and complete reference for most current antibody techniques.
Specific Reference
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Product Type
GFP and Epitopes;
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Product Type
GFP and Epitopes;
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Secondary Antibodies;
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Conjugation Reference
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Conjugation Chemistry