Immunofluorescence analysis of CUL-2 was performed using 70% confluent log phase Caco-2 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 ABfinity™ CUL-2 (50H17L12) Recombinant Rabbit Monoclonal Antibody (700179) 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, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 733-745 of Q13617.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||0.5-1 ug/test|
|Immunocytochemistry (ICC)||8-12 ug/ml|
|Immunofluorescence (IF)||8-12 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||4-6 ug/ml|
|Western Blot (WB)||3-5 ug/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.
This antibody is predicted to react with bovine, canine, chimpanzee, equine, orangutan, primate, Rhesus monkey and Xenopus based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
In yeast, proteolysis of G1 cell cycle regulatory proteins is controlled by an ubiquitin ligase formed by three subunits: Cdc53 (also known as CulA), Skp1 and one of many F-box proteins. A family of human cullin genes homologous to the S. cerevisiae cdc53 gene has been identified (cul1, 2, 3, 4a, 4b, and 5). In humans Cul2 is part of the ECS (Elongin B/C-Cul2/Cul5-SOCS-box protein) complex which contains the Cullin-RBX module. Human Cul2 does not associate with Skp1 and Skp2. Instead it forms a complex with an inactive transcriptional elongation complex, SIII, formed by three subunits: Elongin C, Elongin B and VHL. The Cul2-Rbx1 module specifically interacts with the VHL box. Intriguingly, Elongin C shows homology with Skp1 and Elongin B with ubiquitin. In its active form, SIII does not contain VHL but rather contains an F box protein, Elongin A.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.