Immunofluorescence analysis of SMT3/ SUMO3 was done on 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with SMT3 / SUMO 3 (1H9L17), ABfinity™ Rabbit Monoclonal Antibody (700186) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit Secondary Antibody, Alexa Fluor® 488 conjugate at 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 is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 91-103 of P55854.|
|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)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||4-6 ug/ml|
|Immunofluorescence (IF)||1-5 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||4-6 ug/ml|
|Western Blot (WB)||1:1000|
* 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 1 publications below|
This antibody is predicted to react with chimpanzee and Rhesus monkey 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.
Small ubiquitin-related modifier proteins are members of the ubiquitin-like protein family. SUMO-3 regulates Amyloid beta generation and may be critical in the onset or progression of Alzheimer's disease. SUMO-2/3 forms poly-. SUMO-1, 2, and 3 proteins localize to the nuclear membrane, nuclear bodies, and cytoplasm, respectively. SUMOylation is essential for mammalian cell cycle regulation, SUMO-2/3 localizes to centromeres and condensed chromosomes where binds CENP-E.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Preserving protein profiles in tissue samples: differing outcomes with and without heat stabilization.
700186 was used in western blot to study the effects of different post-mortem sample preparation techniques on the in vivo proteome of mouse hippocampus and cortex
|Ahmed MM,Gardiner KJ||Journal of neuroscience methods (196:99)||2011|