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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a
  1. Products /
  2. Primary Antibodies /
  3. Protein Folding and Stability Antibody Sampler Kit

Protein Folding and Stability Antibody Sampler Kit #4889

    Product Information

    Kit Usage Information

    Protocols

    Product Description

    This sampler kit provides an economical means to investigate protein folding and stability. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

    Specificity / Sensitivity

    Skp1 Antibody detects endogenous levels of total Skp1 protein. Skp2 (D3G5) XP® Rabbit mAb recognizes endogenous levels of total Skp2 protein. This antibody is predicted to cross-react with Skp2α and Skp2ß. ISG15 (22D2) Rabbit mAb detects endogenous levels of the uncleaved precursor form of ISG15 protein. This antibody does not recognize the activated (cleaved) or conjugated forms of ISG15. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3 and NEDD8. NEDD8 (19E3) Rabbit mAb detects endogenous levels of both free and conjugated NEDD8 protein. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3 and ISG15. Ubiquitin (P4D1) Mouse mAb detects ubiquitin, polyubiquitin and ubiquitinated proteins. UBC3 Antibody detects endogenous levels of total UBC3 and UBC3B protein. SUMO-1 Antibody detects recombinant SUMO-1 and endogenous levels of sumoylated proteins (e.g. SUMO-1-RanGAP, 90kD). SUMO-2/3 Antibody detects endogenous levels of SUMO-2/3, but does not cross-react with recombinant SUMO-1.

    Source / Purification

    Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to human SUMO-1 (#4930), UBC3 (#4997), and Skp1 (#2156). Polyclonal antibodies are purified by protein A and peptide affinity chromatography. SUMO-2/3 (18H8) Rabbit mAb (#4971) is produced by immunizing rabbits with a synthetic peptide from the amino terminus of human SUMO-3. Ubiquitin (P4D1) Mouse monoclonal antibody (#3936) is produced by immunizing mice with 1-76 full-length ubiquitin of bovine origin. ISG15 (22D2) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to amino acids at the amino terminus of human ISG15 protein. NEDD8 (19E3) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to amino acids at the amino terminus of human NEDD8 protein. Skp2 (D3G5) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Skp2 protein.

    Background

    The small regulatory protein ubiquitin is often covalently linked to many cellular proteins, labeling these targeted proteins for proteasome-mediated degradation. Ubiquitin is first activated by forming a thiolester complex with the E1 activation component. Activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then to an E3 ubiquitin ligase for final delivery to the ε-NH2 of the target protein lysine residue (1). The ubiquitin-proteasome pathway has been implicated in a wide range of both normal biological processes and diseases (2,3).
    The ubiquitin-like protein family contains three small ubiquitin-related modifier proteins (SUMO-1, -2 and -3), neural precursor cell-expressed developmentally down-regulated protein 8 (NEDD8) and interferon-stimulated 15 kDa protein (ISG15) (4-6). Their covalent attachment to target proteins is a reversible, multi-step process that is analogous to protein ubiquitination. Mature molecules are linked to the activating enzyme E1, conjugated to E2 and ligated to the target proteins by E3 (7-10). Ubiquitin is the predominant regulator for the degradation of a wide range of target proteins (8) while SUMO, NEDD8 and ISG15 modify a limited set of substrates to regulate various other biological processes (4, 11-18).
    During ubiquitination, the combinatorial interaction of different E2 and E3 proteins produces variable substrate specificity (4). UBC3 and UBC3B are E2 ubiquitin-carrier proteins (19, 20). The SCF (Skp1/CUL1/F-box) E3 ubiquitin ligase protein complex is composed of three protein components, including the S phase kinase associated protein 1 (Skp1), Cullin homolog 1 (CUL1) and the Skp2 F-box protein (21-23).
    1. Ciechanover, A. (1998) EMBO J. 17, 7151-7160.
    2. Bernardi, R. et al. (2000) Oncogene 19, 2447-2454.
    3. Jesenberger, V. and Jentsch, S. (2002) Nat. Rev. Mol. Cell Biol. 3, 112-121.
    4. Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem. Sci. 28, 321-328.
    5. Chiba, T. and Tanaka, K. (2004) Curr. Protein Pept. Sci. 5, 177-184.
    6. Ritchie, K.J. and Zhang, D.E. (2004) Semin. Cell Dev. Biol. 15, 237-246.
    7. Kim, K.I. et al. (2002) J. Cell Physiol. 191, 257-268.
    8. Osaka, F. et al. (1998) Genes Dev. 12, 2263-2268.
    9. Loeb, K.R. and Haas, A.L. (1992) J. Biol. Chem. 267, 7806-7813.
    10. Zhao, C. et al. (2005) Proc. Natl. Acad. Sci. USA 102, 10200-10205.
    11. Matunis, M.J. et al. (1996) J. Cell Biol. 135, 1457-1470.
    12. Duprez, E. et al. (1999) J. Cell Sci. 112 ( Pt 3), 381-393.
    13. Gostissa, M. et al. (1999) EMBO J. 18, 6462-6471.
    14. Rodriguez, M.S. et al. (1999) EMBO J. 18, 6455-6461.
    15. Desterro, J.M. et al. (1998) Mol. Cell 2, 233-239.
    16. Stickle, N.H. et al. (2004) Mol. Cell. Biol. 24, 3251-3261.
    17. Xirodimas, D.P. et al. (2004) Cell 118, 83-97.
    18. Hamerman, J.A. et al. (2002) J. Immunol. 168, 2415-2423.
    19. Semplici, F. et al. (2002) Oncogene 21, 3978-3987.
    20. Pagano, M. et al. (1995) Science 269, 682-685.
    21. Yu, Z.K. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 11324-11329.
    22. Pagano, M. (2004) Mol. Cell 14, 414-416.
    23. Reed, S.I. (2003) Nat. Rev. Mol. Cell Biol. 4, 855-864.

    Database Links

    UniProt ID: Q15843 , P55854 , Q712K3 , P61956 , P63208 , P05161 , P63165 , P62987 , P0CG48 , Q13309 , P0CG47 , P62979 , P49427


    Entrez-Gene Id: 4738 , 6612 , 54926 , 6613 , 6500 , 9636 , 7341 , 7311 , 7316 , 6502 , 7314 , 6233 , 997


    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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