Immunofluorescent analysis of Survivin was performed on 70% confluent log phase HeLa 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 ABfinity™ Survivin recombinant rabbit monoclonal antibody (Product # 700387) at a dilution of 1:1000 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381) Panel d is a merged image showing nuclear localization and panel e is a control without primary antibody. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Bovine, Hamster, Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein corresponding to amino acids 1-142.|
|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)||1:100-1:500|
|Western Blot (WB)||1:500-1:5000|
* 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 and hamster 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.
Survivin is the smallest member of the Inhibitors of Apoptosis Protein (IAP) gene family and is also known as IAP4. The IAPs are involved in multiple cell functions such as cell signaling, cell division and metabolism. It exhibits differential expression in nearly all human cancers, but not in most normal tissues. Survivin has been shown to express in a cell cycle-dependent manner in the G2/M cell cycle and associates with the mitotic apparatus. The expression of surviving has been shown to be down regulated by expression of wild-type p53.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.