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Immunofluorescent analysis of phospho-AKT pSer473 (green) in HeLa cells either serum starved overnight (left panel) or treated with 10% FBS (right panel) for 30 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a phospho-AKT pSer473 monoclonal antibody (Product # 44-621G) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antibody was produced against a chemically synthesized phosphopeptide derived from the region of human Akt that contains serine 473.|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||1:200-1:2000|
* 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 product contains enough material for 10 mini-blots.
The immunogenic sequence is conserved among multiple species including human, mouse and rat in all three isoforms: Akt1, Akt2 and Akt3.
Recommended positive controls include NIH-3T3 cells stimulated with PDGF and 3T3-L1 adipocytes stimulated with OSM and IFN-gamma.
AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A high-fish-oil diet prevents adiposity and modulates white adipose tissue inflammation pathways in mice.
44-621G was used in western blot to study the regulatory effect of a high-fish-oil diet on rodent white adipose tissue inflammation pathways
|Bargut TC,Mandarim-de-Lacerda CA,Aguila MB||The Journal of nutritional biochemistry (26:960)||2015|
p53 attenuates AKT signaling by modulating membrane phospholipid composition.
44-621G was used in western blot to report a role for p53 in regulating the membrane phospholipids composition.
|Rueda-Rincon N,Bloch K,Derua R,Vyas R,Harms A,Hankemeier T,Khan NA,Dehairs J,Bagadi M,Binda MM,Waelkens E,Marine JC,Swinnen JV||Oncotarget (6:21240)||2015|
Ganitumab (AMG 479) inhibits IGF-II-dependent ovarian cancer growth and potentiates platinum-based chemotherapy.
44-621G was used in western blot to assess the therapeutic potential of ganitumab for the treatment of ovarian cancer.
|Beltran PJ,Calzone FJ,Mitchell P,Chung YA,Cajulis E,Moody G,Belmontes B,Li CM,Vonderfecht S,Velculescu VE,Yang G,Qi J,Slamon DJ,Konecny GE||Clinical cancer research : an official journal of the American Association for Cancer Research (20:2947)||2014|
PLC-γ and PI3K link cytokines to ERK activation in hematopoietic cells with normal and oncogenic Kras.
44-621G was used in western blot to demonstrate that phospholipase C-γ, PI3K, and their generated second messengers link activated cytokine receptors to Ras and ERK signaling in differentiated bone marrow cells and leukemia stem cells.
|Diaz-Flores E,Goldschmidt H,Depeille P,Ng V,Akutagawa J,Krisman K,Crone M,Burgess MR,Williams O,Houseman B,Shokat K,Sampath D,Bollag G,Roose JP,Braun BS,Shannon K||Science signaling (6:null)||2013|
Noncovalent wild-type-sparing inhibitors of EGFR T790M.
44-621G was used in western blot to identify indolocarbazole compounds as potentially effective EGFR inhibitors.
|Lee HJ,Schaefer G,Heffron TP,Shao L,Ye X,Sideris S,Malek S,Chan E,Merchant M,La H,Ubhayakar S,Yauch RL,Pirazzoli V,Politi K,Settleman J||Cancer discovery (3:168)||2013|
Type I IFN receptor and the B cell antigen receptor regulate TLR7 responses via distinct molecular mechanisms.
44-621G was used in western blot to test if BCR-mediated effects on TLR7 tolerance are mediated by type I IFN receptor signals.
|Poovassery JS,Bishop GA||Journal of immunology (Baltimore, Md. : 1950) (189:1757)||2012|
Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors.
44-621G was used in western blot to study the effect of receptor tyrosine kinases ligands on cancer resistance.
|Wilson TR,Fridlyand J,Yan Y,Penuel E,Burton L,Chan E,Peng J,Lin E,Wang Y,Sosman J,Ribas A,Li J,Moffat J,Sutherlin DP,Koeppen H,Merchant M,Neve R,Settleman J||Nature (487:505)||2012|
|The role of SHIP in the development and activation of mouse mucosal and connective tissue mast cells.||Ruschmann J,Antignano F,Lam V,Snyder K,Kim C,Essak M,Zhang A,Lin AH,Mali RS,Kapur R,Krystal G||Journal of immunology (Baltimore, Md. : 1950) (188:3839)||2012|
Protein phosphatase magnesium dependent 1A (PPM1A) plays a role in the differentiation and survival processes of nerve cells.
44-621G was used in western blot to study the role of PPMA in the survival and differentiation processes of PC6-3 cells.
|Shohat M,Ben-Meir D,Lavi S||PloS one (7:null)||2012|
Neuregulin-1-mediated autocrine signaling underlies sensitivity to HER2 kinase inhibitors in a subset of human cancers.
44-621G was used in western blot to suggest that lapatinib may benefit patients with NRG-driven tumors lacking HER2 amplification.
|Wilson TR,Lee DY,Berry L,Shames DS,Settleman J||Cancer cell (20:158)||2011|
||The role of SHIP in the development and activation of mouse mucosal and connective tissue mast cells.||Ruschmann J,Antignano F,Lam V,Snyder K,Kim C,Essak M,Zhang A,Lin AH,Mali RS,Kapur R,Krystal G||Journal of immunology (Baltimore, Md. : 1950) (188:3839)||2012|
AKT; AKT1 kinase; AKT1m; Akt1m protein; C-AKT; EC 188.8.131.52; kinase Akt1; PKB; PKB alpha; PKB-alpha; Protein kinase B; Protein kinase B alpha; protein kinase B-alpha; Proto-oncogene c-Akt; RAC; rac protein kinase alpha; RAC-alpha serine/threonine kinase; RAC-alpha serine/threonine-protein kinase; RAC-PK-alpha; related to A and C kinases; serine-threonine protein kinase; Thymoma viral proto-oncogene; v-akt murine thymoma viral oncogene homolog 1; v-akt murine thymoma viral oncogene-like protein 1
AKT; AKT1; CWS6; PKB; PKB-ALPHA; PKB/Akt; PKBalpha; PRKBA; RAC; RAC-ALPHA