Product Pathways - Protein Folding
Protein Folding and Stability Antibody Sampler Kit #4889
|4889S||1 Kit (8 x 40 µl)||---||In Stock||---|
Already purchased this product? Write a Review.
|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Skp1 Antibody #2156||40 µl||W, IF-IC, F||H, M, R||19||Rabbit|
|Skp2 (D3G5) XP® Rabbit mAb #2652||40 µl||W, IP, IHC-P, IF-IC||H, Mk||48||Rabbit IgG|
|ISG15 (22D2) Rabbit mAb #2758||40 µl||W, IP||H, Hr||15||Rabbit IgG|
|NEDD8 (19E3) Rabbit mAb #2754||40 µl||W, IP, IHC-P, F||H, M, R, Mk||X, Z, B||9||Rabbit IgG|
|Ubiquitin (P4D1) Mouse mAb #3936||40 µl||W, IHC-P||All||Mouse IgG1|
|UBC3 Antibody #4997||40 µl||W, F||H, M, R||32||Rabbit|
|SUMO-1 Antibody #4930||40 µl||W, IP, IHC-P, IF-IC||H, M, R, Mk||Rabbit|
|SUMO-2/3 (18H8) Rabbit mAb #4971||40 µl||W, IF-IC||H, R||M||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||Horse|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Hr=Horse, All=All Species Expected
Western blot analysis of extracts from PC12 and NIH/3T3 cells using Skp1 Antibody #2156.
Western blot analysis of extracts from various cell lines using Skp2 (D3G5) XP® Rabbit mAb #2652.
Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb #2754.
Western blot analysis of lysates from HeLa and A549 cells, untreated (-) or treated with IFN-α (1000 U/mL) (+) for 24 hours, using ISG15 (22D2) Rabbit mAb #2758.
Western blot analysis of extracts from 293 and HeLa cells, untreated or MG-132 (a 26S proteosome inhibitor) treated (50 µM for 90 minutes), using Ubiquitin (P4D1) Mouse mAb #3936.
Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa) and extracts from CAD, HeLa and PC-12 cells using SUMO-1 Antibody #4930.
Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa), recombinant SUMO-2, recombinant SUMO-3, and extracts from C6 and PC12 cells, using SUMO-2/3 (18H8) Rabbit mAb #4971 (upper) and SUMO-1 Antibody #4930 (lower).
This sampler kit provides an economical means to investigate protein folding and stability. The kit contains primary and secondary antibodies to perform four 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.
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).
- Ciechanover, A. (1998) EMBO J. 17, 7151-7160.
- Bernardi, R. et al. (2000) Oncogene 19, 2447-2454.
- Jesenberger, V. and Jentsch, S. (2002) Nat. Rev. Mol. Cell Biol. 3, 112-121.
- Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem. Sci. 28, 321-328.
- Chiba, T. and Tanaka, K. (2004) Curr. Protein Pept. Sci. 5, 177-184.
- Ritchie, K.J. and Zhang, D.E. (2004) Semin. Cell Dev. Biol. 15, 237-246.
- Kim, K.I. et al. (2002) J. Cell Physiol. 191, 257-268.
- Osaka, F. et al. (1998) Genes Dev. 12, 2263-2268.
- Loeb, K.R. and Haas, A.L. (1992) J. Biol. Chem. 267, 7806-7813.
- Zhao, C. et al. (2005) Proc. Natl. Acad. Sci. USA 102, 10200-10205.
- Matunis, M.J. et al. (1996) J. Cell Biol. 135, 1457-1470.
- Duprez, E. et al. (1999) J. Cell Sci. 112 ( Pt 3), 381-393.
- Gostissa, M. et al. (1999) EMBO J. 18, 6462-6471.
- Rodriguez, M.S. et al. (1999) EMBO J. 18, 6455-6461.
- Desterro, J.M. et al. (1998) Mol. Cell 2, 233-239.
- Stickle, N.H. et al. (2004) Mol. Cell. Biol. 24, 3251-3261.
- Xirodimas, D.P. et al. (2004) Cell 118, 83-97.
- Hamerman, J.A. et al. (2002) J. Immunol. 168, 2415-2423.
- Semplici, F. et al. (2002) Oncogene 21, 3978-3987.
- Pagano, M. et al. (1995) Science 269, 682-685.
- Yu, Z.K. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 11324-11329.
- Pagano, M. (2004) Mol. Cell 14, 414-416.
- Reed, S.I. (2003) Nat. Rev. Mol. Cell Biol. 4, 855-864.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 4995 CUL1 Antibody
- 2754 NEDD8 (19E3) Rabbit mAb
- 4996 UBC3B Antibody
- 2738 VHL Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 2156 Skp1 Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.