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Goat TrueBlot® Western Blot Kit

Mouse Monoclonal eB270
NCI Collaboration
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Goat TrueBlot® IP / Western Blot:  Jurkat cell lysate (0.5 ml of 1x10e7 cells/ml) was incubated with goat anti-human Stat1 and immunoprecipitated using Protein G. Precipitate from 5x10e5 cells was subjected to electrophoresis, transferred to a PVDF membrane, and Western blotted with anti-Stat1 using Goat TrueBlot®: Anti-Goat IgG HRP (lane 1) and conventional HRP-conjugated anti-goat polyclonal antibody (lane 2).
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Number:
88-8884-31
Size:
1 Kit
Price:
$259.00
Shipping:
$20.00
Handling:
$20.00
Availability:
Ships next business day.

Goat TrueBlot® Western Blot Kit Properties

Description
Goat TrueBlot® Western Blot Kit - 88-8884-31
Host
Mouse
Target Species
Goat
Known Cross Reactivity
Goat
Clonality
Monoclonal eB270
Application
Western Blot : 1:1000
ImmunoPrecipitation: Use with TrueBlot® Anti-Goat Ig IP Beads Cat #( 00-8844-25)
Product Type
Kit
Kit Type
Immunoprecipitation Kit
Physical State
Liquid (sterile filtered)
Shipping Condition
Wet Ice
Format
IgG
Buffer
0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Stabilizer
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol
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Goat TrueBlot® Western Blot Kit Description

Background
The Goat TrueBlot® Western Blot Kit contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples using TrueBlot second step immunoblotting reagents in conjunction with your own primary IP antibody and primary (Goat IgG) Western blotting antibody. TrueBlot technology enables unhindered detection of protein bands of interest which would otherwise be obscured by the presence of reduced and denatured heavy and light chain immunoglobulin in the blot (as detected by the conventional immunoblotting HRP anti-Goat IgG reagent). Goat IgG TrueBlot® is the unique horseradish peroxidase conjugated anti-Goat IgG immunoblotting second step reagent which enables detection of immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your usual HRP anti-Goat IgG immunoblotting second step reagent. It is easy to generate publication-quality IP/WB data with Goat IgG TrueBlot. Goat IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of goat IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Goat 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/immunoblotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions. Goat IgG TrueBlot® may also be used for detection in immunoblotting assays that do not employ immunoprecipitation.
Synonyms
HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting
Storage Condition
Store at 2-8°C except Goat TrueBlot® which should be stored at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Application Note
Goat IgG TrueBlot® is provided as 1000X solution. 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. 18-8814-31 (50µL) will yield enough reagent for 20 blots. Goat IgG TrueBlot®. An HRP-conjugated second step reagent reacting with goat IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 2. Immunoprecipitate should be completely reduced. 3. Milk should be used as the blocking protein for the immunoblot. A sample of Anti-alpha-Tubulin (MOUSE) Monoclonal Antibody Cat #(200-301-880S), has been provided to be used as a loading control at the user’s discretion. Dilutions are to be optimized by the user. The Goat TrueBlot Western Blot Kit components are sufficient for 20-25 miniblots. Components: 1. Goat IgG TrueBlot: 50 µl. 18-8814-31 2. TrueBlot Enhancer Solution: 25 ml 3. TrueBlot Blocker: 10 g 4. TrueBlot Assay Buffer: 30 ml. 20X 5. TrueBlot Substrate A: 12.5 ml 6. TrueBlot Substrate B: 12.5 ml 7. Western Blot Incubation Tray 8. Anti-alpha-Tubulin (MOUSE) Monoclonal Antibody - 200-301-880S
Purity/Specifity
Goat TrueBlot® Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum. Reactivity is observed against native Goat 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 5199, Limerick, 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
WB
Reacts With
Goat
Catalog Number
18-8814-31

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Goat
Catalog Number
18-8814-33

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Sheep
Catalog Number
18-8815-31

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Sheep
Catalog Number
18-8815-33

Product Label

 

Description
HRP TrueBlot® ULTRA for IP/WB
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry