Immunofluorescence analysis of AKT pan was done on 70% confluent log phase U2OS 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 AKT pan Rabbit Polyclonal Antibody (44609G) at 2 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (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 (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the carboxy-terminal region of human Akt1.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-3 ug/ml|
|Immunofluorescence (IF)||1-3 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||0.5-2 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.
AKT also known as protein kinase B (PKB) or RAS-alpha, is an ubiquitous serine/threonine kinase that plays an important role in diverse biological responses such as regulation of metabolism, cell survival and growth by phosphorylating multiple proteins. This protein kinase is activated by insulin, PI3K, IGF1 and various other growth and survival factors. Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. The AKT pathway is a major target for cancer drug discovery.
IP-MS enrichment of AKT1 (LFQ intensity): AKT1 was enriched 621-fold from HCT116 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and AKT1 antibody (Part No. 44-609G). See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Pooled screening for antiproliferative inhibitors of protein-protein interactions.
44-609G was used in western blot to assess antiproliferative inhibitors of protein-protein interactions by pooled screening
|Nim S,Jeon J,Corbi-Verge C,Seo MH,Ivarsson Y,Moffat J,Tarasova N,Kim PM||Nature chemical biology (12:275)||2016|
A high-fish-oil diet prevents adiposity and modulates white adipose tissue inflammation pathways in mice.
44-609G 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.
44-609G 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|
GPR30 activation is neither necessary nor sufficient for acute neuroprotection by 17ß-estradiol after an ischemic injury in organotypic hippocampal slice cultures.
44-609G was used in western blot to examine the role of GPR30 in 7β-estradiol-mediated neuroprotection after an ischemic injury in an organotypic hippocampal slice culture model.
|Lamprecht MR,Morrison B||Brain research (1563:131)||2014|
|Mouse||Not Cited||Signaling in sperm: toward a molecular understanding of the acquisition of sperm motility in the mouse epididymis.||Vadnais ML,Aghajanian HK,Lin A,Gerton GL||Biology of reproduction (89:null)||2013|
Adverse effects of vitamin D deficiency on the Pi3k/Akt pathway and pancreatic islet morphology in diet-induced obese mice.
44-609G was used in western blot to examine the impact of vitamin D deficiency on insulin resistance and abnormal glucose homeostasis in obesity.
|Borges CC,Salles AF,Bringhenti I,Souza-Mello V,Mandarim-de-Lacerda CA,Aguila MB||Molecular nutrition and food research (60:346)||2016|
Effect of whole-body vibration and insulin-like growth factor-I on muscle paralysis-induced bone degeneration after botulinum toxin injection in mice.
44-609G was used in immunohistochemistry to test if whole-body vibration and insulin-like growth factor-I treatment counteract paralysis-induced bone degradation following botulinum toxin A injections via protein kinase B signaling.
|Niehoff A,Lechner P,Ratiu O,Reuter S,Hamann N,Brüggemann GP,Schönau E,Bloch W,Beccard R||Calcified tissue international (94:373)||2014|