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Fluorescent Western Blot Kit: MaxTag™ for IRDYE® Immunoblotting for use with Rabbit and Mouse Primary Antibodies (Rb800/Ms700) - KFA011
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Blocking Buffer for Fluorescent Western Blotting - MB-070
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Kits  >  Fluorescent Western Blot Kits
 

Fluorescent Western Blot Kit: MaxTag™ for IRDYE® Immunoblotting

Code

KFA012
Size

1 Each
Price

$245.00
Availability

Ships next business day.
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Phone: 800.656.7625
E-mail: orders@rockland-inc.com
Fax: 610.367.7825
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Sensitivity of Fluorescent Western Blotting Detection-Fluorescent Western blots show detection of a dilution series of alpha-1-anti-trypsin (a1AT ) protein, detected with rabbit anti-a1AT antibody and IRDye™800 conjugated anti-Rabbit IgG secondary antibody.  The antigen was loaded using a 2-fold dilution series starting at 2 ng.  The amount loaded into each lane is marked at the top of the western blot.  Both the Blocking Buffer and Washing Buffer provided in this kit were used to process this blot.  Primary antibody was diluted to 5 µg/ml and reacted overnight at +4°C.   In Panel A the image is shown in black and white.  Panel B shows the same image displayed in color as green.  As little as 8 pg of protein can easily be detected using this system.
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Synonyms: Duplex Western Blotting, IRDYE™ Kit for immunoblotting, IRDYE™800 kit, IRDYE™700 kit

Fluorescent Western Blot Kit: MaxTag™ for IRDYE® Immunoblotting Properties

Description
Fluorescent Western Blot Kit: MaxTag™ for IRDYE® Immunoblotting for use with Mouse and Rabbit Primary Antibodies (Ms800/Rb700) - KFA012
Known Cross Reactivity
Anti-Mouse and Anti-Rabbit IgG
Application
Western Blot : 1:5,000
Product Type
Kit
Kit Type
Fluorescent Western Blot Kit
Physical State
n/a
Shipping condition
Wet Ice
Label
Concentration
Sufficient to run approximately 25 immunoblots (10 cm x 10 cm).
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Fluorescent Western Blot Kit: MaxTag™ for IRDYE® Immunoblotting Description

Background
Use Rockland's MaxTag™ Kit for Duplex IRDYE® Immunoblotting for the detection of Rabbit and Mouse primary antibodies using infrared-labeled antibodies to form an antigen-antibody-antibody complex that is immobilized on a membrane (immunoblot) and detected using a two channel infrared scanner such as LI-COR's Odyssey® Infrared Imaging System. This Kit is useful for both “western blotting” and “dot blotting” methods and can be used for single (1-color) or duplex (2-color) antibody detection (see Figure 3). Nitrocellulose membrane formats are preferred (see sections XI and XII for more details). If you require additional assistance please call or e-mail our technical service representatives at 800-656-7625 or tech@rockland-inc.com.
Storage Condition
See kit insert for complete instructions.
Application Note
Infrared-labeled antibodies and controls provided in this kit are sufficient to run approximately 25 immunoblots each measuring 10 cm x 10 cm. The amount of antibody supplied when diluted as recommended in our protocol will yield 1000 ml of working solution. Adjustments in volumes for larger or smaller blots will effect the number of blots detected. Blocking and washing buffers are provided in limited quantities so that the user can immediately begin to process immunoblots. These buffers can be re-ordered in larger quantities for economical use (see ''Replacement Parts'').
Purity/Specifity
This kit allows for the individual (1-color) or simultaneous (2-color) detection of primary rabbit antibodies using an IRDye®800 conjugated secondary antibody (green) and primary mouse antibodies using an IRDye®700DX conjugated secondary antibody (red). The user is required to provide all primary antibodies. After protein separation and transfer, the membrane is blocked to prevent non-specific protein binding using a specially formulated blocking buffer for infrared fluorescent immunoblot detection. After washing the membrane, user-provided unlabeled primary antibodies are bound to the antigen. Additional washings are followed by the addition of IRDye®800 and IRDye®700DX conjugated secondary antibodies (see Figure 1). Final washing removes excess reagents and the membrane is further prepared for simultaneous scanning at two wavelengths to detect antibody complexes using a two channel infrared scanner such as LI-COR's Odyssey® Infrared Imaging System. The labeled antibodies are detected directly on the membrane, eliminating film, darkrooms, and messy substrates, while preserving high sensitivity (see Figure 2).
Disclaimer Note-General
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 326, Gilbertsville, Pennsylvania, USA.
General Reference
Specific Reference
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Product Type
Kits;
Applications
WB
Reacts With
Anti-Rabbit and Anti-Mouse IgG
Catalog Number
KFA011

Product Type
Supporting Reagents;
Applications
WB
Catalog Number
MB-070

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry