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Immunofluorescent analysis of p70S6K (pT389) was performed on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% of Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ p70S6K (pT389) recombinant rabbit monoclonal antibody (Product # 701064) at a dilution of 1:500 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 cytoplasmic and nuclear localization. Panel e shows competition with p70S6K (pT389) peptide. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphopeptide corresponding to amino acids 384–394 of human p70 ribosomal protein S6 kinase|
|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.
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.
p70 Ribosomal Protein S6 Kinase (p70S6K) is a 70 kDa member of the ribosomal S6 kinase (RSK) family of serine/threonine kinases. It is predominantly localized in the cytoplasm, and is essential in growth factors regulated cell proliferation, pathways involving cell motility, such as metastases, the immune response, and tissue repair. p70S6K acts downstream of phosphoinositide (PI) 3-kinase, and its main physiological target is the S6 ribosomal protein, which is involved in upregulation of protein synthesis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
alpha 1; alpha 2; p70 S6 kinase; p70 S6 kinase alpha; p70 S6 kinase, alpha; p70 S6K-alpha; p70 S6KA; p70-S6K 1; P70S6K1; ribosomal protein S6 kinase beta-1; ribosomal protein S6 kinase I; ribosomal protein S6 kinase, 70kDa, polypeptide 1; serine/threonine kinase 14 alpha; serine/threonine-protein kinase 14A
p70 S6KA; p70(S6K)-alpha; p70-alpha; p70-S6K; PS6K; RPS6KB1; S6K; S6K-beta-1; S6K1; STK14A