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Western blot analysis was performed on whole cell, tissue extracts (30 ug lysate) of HEL 92.1.7 (Lane 1), MDA-MB-231 (Lane 2), NTERA-2 (Lane 3), MCF-7 (Lane 4), A549 (Lane 5), Caco-2 (Lane 6), Mouse Testis (Lane 7), Rat Testis (Lane 8) and Mouse pancreas (Lane 9). The blots were probed with Anti- CUL1 Mouse Monoclonal Antibody (Product# 322400, 1-2 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 ug/ml, 1:2500 dilution). A ~ 89 kDa band corresponding to CUL1 was observed across cell lines tested. Additionally ~25 kDa band was observed across mouse species. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with Pierce™ Power Blotter System (Product # 22834). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Insect, Virus, Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||A synthetic peptide corresponding to an 11 amino acid sequence at the C-terminus of the human CUL-1 protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||5-10 ug|
|Western Blot (WB)||1-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
CUL-1 Antibody (2H4C9), Prod# 32-2400 specifically detects neddylated and unmodified Cullin 1 in human cells.
Cullin 1 is a member of the family of human cullin genes (CUL 1, 2, 3, 4a, 4b, and 5) homologous to the S. cerevisiae cdc 53 gene. CUL 1 forms a complex with human p19 skp1 and F box protein p45 skp2, both of which, together with cdc 53 (known as CUL A), as three subunits form a ubiquitin ligase controlling proteolysis of G1 cell cycle regulatory proteins. The CUL 1/p19 skp1/p45 skp2complex is thought to play a role in the ubiquitination of G1 regulatory proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Ubiquitin-conjugating enzyme Cdc34 and ubiquitin ligase Skp1-cullin-F-box ligase (SCF) interact through multiple conformations.
32-2400 was used in western blot to study the interaction between ubiquitin-conjugating enzyme Cdc34 and ubiquitin ligase Skp1-cullin-F-box ligase.
|Sandoval D,Hill S,Ziemba A,Lewis S,Kuhlman B,Kleiger G||The Journal of biological chemistry (290:1106)||2015|
F-box protein FBXL16 binds PP2A-B55α and regulates differentiation of embryonic stem cells along the FLK1+ lineage.
32-2400 was used in western blot to study the interaction between F-box protein FBXL16 and PP2A-B55alpha and its effect on the differentiation of embryonic stem cells.
|Honarpour N,Rose CM,Brumbaugh J,Anderson J,Graham RL,Sweredoski MJ,Hess S,Coon JJ,Deshaies RJ||Molecular & cellular proteomics : MCP (13:780)||2014|
|Insect||Not Cited||UBXN7 docks on neddylated cullin complexes using its UIM motif and causes HIF1α accumulation.||Bandau S,Knebel A,Gage ZO,Wood NT,Alexandru G||BMC biology (10:null)||2012|
Early insights into the function of KIAA1199, a markedly overexpressed protein in human colorectal tumors.
32-2400 was used in immunoprecipitation to examine the localization and function of KIAA99 in human colorectal tumors.
|Tiwari A,Schneider M,Fiorino A,Haider R,Okoniewski MJ,Roschitzki B,Uzozie A,Menigatti M,Jiricny J,Marra G||PloS one (8:null)||2013|
|Virus||Not Cited||Cullin-3 regulates late endosome maturation.||Huotari J,Meyer-Schaller N,Hubner M,Stauffer S,Katheder N,Horvath P,Mancini R,Helenius A,Peter M||Proceedings of the National Academy of Sciences of the United States of America (109:823)||2012|