Goat TrueBlot®: Anti-Goat IgG HRP

RUO: For Research Use Only

goat IgG antibody, Horseradish Peroxidase (HRP) conjugate

Jurkat cell lysate (0.5 ml of 1x10⁷ cells/ml) was incubated with goat anti-human Stat1 and immunoprecipitated using Protein G. Precipitate from 5x10⁵ cells was subjected to electrophoresis, transferred to a PVDF membrane, and immunoblotted with anti-Stat1 using Goat TrueBlot®: Anti-Goat IgG HRP (lane 1) and conventional HRP-conjugated anti-goat polyclonal antibody (lane 2).

Contents: Goat TrueBlot®: Anti-Goat IgG HRP
Catalog Number: 18-8814
Formulation: Phosphate buffer pH 7.2, protein stabilizers, 50% glycerol
Storage Conditions: Store at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.

Product Options
Cat. No.	Size	Price	Add Qty to Cart
18-8814-31	50 ul (1000X)	To view relevant pricing, please select your country and resume browsing.
18-8814-33	200 ul (1000X)

Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.

Research Related Products
Cat. No.	Format
00-8844	TrueBlot® Anti-Goat Ig IP Beads
88-1688	Rabbit TrueBlot® Set (with IP beads)

Available Formats
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications	Regulatory Status
18-8814	Goat TrueBlot®: Anti-Goat IgG HRP	N/A	N/A	WB	RUO
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.

Description

Goat IgG TrueBlot® is a unique horseradish peroxidase conjugated anti-goat IgG immunoblotting (second step) reagent. Goat 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/WB data with Goat IgG TrueBlot, simply substitute the conventional HRP anti-goat IgG blotting reagent with Goat IgG TrueBlot and follow the prescribed protocol for sample preparation and immunoblotting.

Note that there are two key procedural considerations:
1. Immunoprecipitate should be completely reduced.
2. Milk should be used as the blocking protein for the immunoblot.

Applications Reported

For research use only, not for diagnostic or therapeutic use. Goat TrueBlot has been reported for use in immunoblotting (WB).

Applications Tested

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.

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.

References

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-κB Kinase complex, inhibits signaling to NF-κB by the Tumor Necrosis Factor superfamily of receptors, and inhibits NF-κB 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)

Okoshi Y, Tahara-Hanaoka S, Nakahashi C, Honda S, Miyamoto A, Kojima H, Nagasawa T, Shibuya K, Shibuya A. 2005. Requirement of the tyrosines at residues 258 and 270 of MAIR-I in inhibitory effect on degranulation from basophilic leukemia RBL-2H3. Int Immunol. 17(1):65-72. (Mouse IgG TrueBlot, PubMed)

Murray, J., M.F. Marusich, R.A. Capaldi, and R. Aggeler. 2004. Focused proteomics: Monoclonal antibody-based isolation of the oxidative phosphorylation machinery and detection of phosphoproteins using a fluorescent phosphoprotein gel stain. Electrophoresis. 25:2520-2525. (Mouse IgG TrueBlot, PubMed)

Hamdane, M., A. Bretteville, A. Sambo, K. Schindowski, S. Begard, A. Delacourte, P. Bertrand, and L. Buee. 2005. p25/Cdk5-mediated retinoblastoma phosphorylation is an early event in neuronal cell death. Journal of Cell Science. 118: 1291-1298. (Rabbit and Mouse IgG TrueBlot, PubMed)

Protocol

To view protocol, please download the TDS by clicking the PDF button above.

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Unless indicated otherwise, all products are For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for further distribution without written consent. Not all products available in all regions.