Immunofluorescence analysis of CUL-3 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CUL-3 Rabbit Polyclonal Antibody (342200) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the N-terminal sequence of human Cullin 3.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
|Western Blot (WB)||See 3 publications below|
Cullin 3 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. Together with cdc 53 (Known as Cul A), as three subunits, they form a ubiquitin ligase-controlling proteolysis of G1 cell cycle regulatory proteins. In contrast to CUL 1, CUL 2 does not associate with p19 skp1 and p45 skp2. Instead, CUL 2 forms a complex with an inactive transcriptional elongation complex, SIII, formed by three subunits: Elongain C, Elongin B and VHL. The function of this association remains unknown.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Mechanism of cullin3 E3 ubiquitin ligase dimerization.
34-2200 was used in western blot to investigate factors that mediate Cul3 interactions.
|Choo YY,Hagen T||PloS one (7:null)||2012|
Neddylation-induced conformational control regulates cullin RING ligase activity in vivo.
34-2200 was used in western blot to study the effect of neddylation on Cul2 and Cul3 structure.
|Boh BK,Smith PG,Hagen T||Journal of molecular biology (409:136)||2011|
|Human||Not Cited||Cullin 3 promotes proteasomal degradation of the topoisomerase I-DNA covalent complex.||Zhang HF,Tomida A,Koshimizu R,Ogiso Y,Lei S,Tsuruo T||Cancer research (64:1114)||2004|