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Secondary Antibodies  >  Fluorescein Conjugated Antibodies
 

Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein

Mouse Monoclonal eB182
Code

18-0216-32
Size

100 µL
Price

$179.00
Availability

Ships next business day.
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Phone: 800.656.7625
E-mail: orders@rockland-inc.com
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Rabbit TrueBlot® IP / Western Blot:

Jurkat cell lysate (0.5 ml of 1x10e7 cells/ml) was incubated with rabbit anti-human Stat1 and immunoprecipitated using Protein G, Protein A and Anti-Rabbit Ig IP Beads. Precipitate from 5x10e5 cells was subjected to electrophoresis, transferred to a PVDF membrane, and Western blotted with anti-Stat1 using Rabbit TrueBlot®: Anti-Rabbit IgG HRP
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Handling:
$20.00
Synonyms: FITC, TrueBlot, FITC TrueBlot ULTRA, Fluorescein TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Rb TrueBlot

Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein Properties

Description
Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein - 18-0216-32
Host
Mouse
Target Species
Rabbit
Known Cross Reactivity
Rabbit
Clonality
Monoclonal eB182
Application
FLISA : User Optimized
IF Microscopy : 1:500 - 1:2,500
FlowCytometry : 1:2,000 - 1:10,000
Western Blot : 1:1000
Immunochemistry: User Optimized
Other Dilution: User Optimized
Product Type
Component
Kit Type
Immunoprecipitation Kit
Physical State
Lyophilized
Shipping condition
Ambient
Format
IgG
FP Value
3.5 moles Fluorescein (FITC) per mole of IgG
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Reconstitution Volume
100 µL
Reconstitution Buffer
Restore with deionized water (or equivalent)
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
10 mg/ml Polyethylene Glycol (PEG-8000)
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Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein Description

Background
Rabbit IgG TrueBlot® is a unique fluorescein conjugated anti-rabbit IgG immunoblotting (second step) reagent. Rabbit IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Fluorescent Western Blot data with Rabbit IgG TrueBlot®, simply substitute the conventional FITC anti-rabbit IgG blotting reagent with Fluorescent Rabbit TrueBlot® Antibody Fluorescein and follow the prescribed protocol for sample preparation and immunoblotting. Rabbit IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of rabbit IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Rabbit IgG TrueBlot® with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/Western blotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions. Fluorescent Rabbit TrueBlot® Antibody Fluorescein may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescein Conjugated Antibodies are designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.
Storage Condition
Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
Fluorescent Rabbit TrueBlot® Antibody Fluorescein is provided as a lyophilized powder. In order to conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mls). If used conservatively at 2.5mls/blot, cat. 18-0216-32 (100µL) will yield enough reagent for 40 blots. Note that there are three key procedural considerations: 1. Protein A or G should not be used for the immunoprecipitation. For immunoprecipitation, anti-rat IgG beads, or anti-rabbit IgG beads should be used for rat or rabbit immunoprecipitating antibodies, respectively. 2. Immunoprecipitate should be completely reduced. 3. Milk should be used as the blocking protein for the immunoblot.
Purity/Specifity
Fluorescent Rabbit TrueBlot® Antibody Fluorescein Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum. Reactivity is observed against native Rabbit IgG by both Western blot and ELISA.
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
Kong, D., L. Xu, Y. Yu, W. Zhu, D.W. Andrews, Y. Yoon, and T.H. Kuo. 2005. Regulation of Ca2+-induced permeability transition by BCL-2 is antagonized by Drp1 and hFis1. Molecular and Cellular Biochemistry. 272: 187-199. (Rabbit IgG TrueBlot, PubMed) DiPerna, G., J. Stack, A.G. Bowie, A. Boyd, G. Kotwal, Z. Zhang, S. Arvikar, E. Latz, K.A. Fitzgerald, and W.L. Marshall. 2004. Poxvirus protein N1L targets the I-kappaB Kinase complex, inhibits signaling to NF-kappaB by the Tumor Necrosis Factor superfamily of receptors, and inhibits NF-kappaB and IRF3 signaling by Toll-like Receptors. J. Biol. Chem. 279: 36570-36578. (Rabbit IgG TrueBlot, PubMed) Zhang, X., Y. Ozawa, H. Lee, Y. Wen, T. Tan, B. Wadzinski, and E. Seto. 2005. Histone deacetylase 3 (HDAC3) activity is regulated by interaction with protein serine/threonine phosphatase 4. Genes & Development. 19: 827-839. (Rabbit IgG TrueBlot, PubMed) Lehtonen, S., E. Lehtonen, K. Kudlicka, H. Holthöfer, and M.G. Farquhar. 2004. Nephrin Forms a Complex with Adherens Junction Proteins and CASK in Podocytes and in Madin-Darby Canine Kidney Cells Expressing Nephrin. Am J Pathol. 165:923-936. (Rabbit IgG TrueBlot, PubMed) Tyagi A, Agarwal C, Harrison G, Glode LM, Agarwal R. 2004. Silibinin causes cell cycle arrest and apoptosis in human bladder transitional cell carcinoma cells by regulating CDKI-CDK-cyclin cascade, and caspase 3 and PARP cleavages. Carcinogenesis. 25: 1711-20. (Mouse IgG TrueBlot, PubMed)
Specific Reference
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Product Type
Secondary Antibodies;
Applications
FLISA, WB, IHC, IF, FC
Reacts With
Mouse
Catalog Number
18-0217-32

Product Type
Secondary Antibodies;
Applications
FLISA, WB, IHC, IF, FC
Reacts With
Rabbit
Catalog Number
18-4416-32

Product Type
Secondary Antibodies;
Applications
FLISA, WB, IF, IP
Reacts With
Rabbit
Catalog Number
18-3216-32

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Rabbit
Catalog Number
18-8816-31

Product Label

 

Description
Fluorescein (FITC)
Color
Green
Conjugation Reference
Modified from The and Feltkamp, Immunology 18; 865, 1970.
Molecular Weight
390
Excitation Wavelength
495
Conjugation Chemistry
Isothiocyanate