Immunofluorescent analysis of Phospho-Rb pSer249/pThr252 in U2OS cells using a Phospho-Rb pSer249/pThr252 recombinant rabbit monoclonal antibody (Product # 701059) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of phosphorylated Rb and Image E is a composite image of cells showing competition with the phospho Rb (pSpT249/252) peptide to inhibit antibody binding.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphopeptide corresponding to amino acids 245–255 of human retinoblastoma 1|
|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)||1-3µg/10^6 cells|
|Immunocytochemistry (ICC)||1-3 ug/ml|
|Immunofluorescence (IF)||1-3 ug/ml|
|Western Blot (WB)||0.1-2 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 non-human primate, mouse and rat 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.
Rb, also known as Retinoblastoma, is a tumor suppressor protein that keeps uncontrolled growth and division in check. Rb belongs to the Pocket Protein family whose members have a pocket for binding proteins. It is nuclear protein which is widely expressed in retinal cells. It is inactivated by phosphorylation and activated by dephosphorylation. Phosphorylation of serine 249 and threonine 252 is catalyzed by Cyclin D-cdk4. In humans, the protein is encoded by the RB1 gene located on the q arm of chromosome 13.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.