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Antibodies

AKT phospho S473 Antibody

Product Sizes
100 ug
200-301-268-100UG
About this Product
SKU:
200-301-268
Additional Names:
AKT1|mouse anti-AKT pS473 Antibody, phospho AKT, RAC-PK-alpha, Protein kinase B, PKB, C-AKT, RAC-alpha serine/threonine-protein kinase, Proto-oncogene c-Akt, AKT1, AKT 1, AKT-1
Application:
Detection/Quantitation, ELISA, Flow Cytometry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride
CE/IVD:
RUO
translate.label.attr.clone:
17F6.B11
Clonality:
Monoclonal
Concentration:
1 mg/ml
Extra Details:
AKT phospho S473 is a component of the PI-3 kinase pathway and is activated by phosphorylation at Ser 473 and Thr 308. AKT is a cytoplasmic protein also known as AKT1, Protein Kinase B (PKB) and rac (related to A and C kinases). AKT is a key regulator of many signal transduction pathways. AKT Exhibits tight control over cell proliferation and cell viability. Overexpression or inappropriate activation of AKT is noted in many types of cancer. AKT mediates many of the downstream events of PI 3-kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI 3-kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis; (ii) promotion of proliferation. Anti-AKT pS473 (MOUSE) Monoclonal Antibody is ideal for investigators involved in Cell Signaling, Cancer, Neuroscience, Signal Transduction research.
Formulation:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Host:
Mouse
Immunogen:
Anti-AKT pS473 (MOUSE) Monoclonal Antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein, followed by hybridoma development.
Isotype:
IgG1, IgG1, kappa
Physical State:
Liquid
Reactivities:
Human, Mouse, Non-human Primate, Rat
Shipping Conditions:
Dry Ice
Specificity:
Anti-AKT pS473 Monoclonal Antibody was purified from concentrated tissue culture supernate by Protein A chromatography. This phospho specific monoclonal antibody is specific for phosphorylated human and mouse AKT protein at S473. A BLAST analysis was used to suggest cross-reactivity with AKT pS473 from human, mouse, rat and chimpanzee sources based on 100% homology with the immunizing sequence. Cr
Storage Conditions:
2-8[o]C Aliquot. Avoid freeze/thaw cycles. Store undiluted.
Supplier:
Rockland Inc
Type:
Antibody: Monoclonal Antibody
Flow Cytometry results of Anti-AKT pS473 (MOUSE) Monoclonal Antibody. The green histogram represents the A431 cells that were stimulated for 15 minutes with 100 ng/mL EGF. The blue histogram shows the untreated A431 cell population, which is bimodal. Both populations were stained with a 1:50 dilution of the Anti-AKT pS473 (MOUSE) Monoclonal Antibody (p/n 200-301-268) for 30 mins at 4°C. The secondary antibody, Anti-MOUSE IgG (H&L) (GOAT) Antibody DyLight™ 488 Conjugated (p/n 610-141-002) was used at a 1:200 dilution for 30 mins at 4°C.
Immunohistochemistry of Mouse anti-AKT pS473 antibody. Tissue: human prostate tissue. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 20 ug/mL for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear and occasionally cytoplasmic. Staining: AKT pS473 as precipitated red signal with hematoxylin purple nuclear counterstain.
Western Blot of Mouse Anti-AKT pS473 antibody. Lane 1: non-phosphorylated AKT in untreated cells. Lane 2: phosphorylated AKT (indicated by arrowhead at ~56 kDa) on PDGF stimulated NIH/3T3 cell lysates. Load: 10 ug per lane. Primary antibody: AKT pS473 antibody at 1:10,000 in TBS with 0.05% Tween-20 with 1% BSA, for 1 h at 4° C. Secondary antibody: HRP conjugated Gt-a-Mouse IgG (p/n 610-103-121) was used at a 1:20,000 dilution for 1 h at 4° C with FemtoMax™ enhanced chemiluminescent reagent (p/n FEMTOMAX-100). Other band(s): none.
Immunofluorescence Microscopy of Mouse Anti-AKTpS473 antibody using STED nanoscopy to evaluate AKT activation and migration. Tissue: A431 cells. Antigen retrieval: Panel A: serum starved,unstimulated cells. Panel B: serum starved, EGF stimulated for 15 mins. A massive increase in AKT-pS473 activation, as measured by intensity signal, peaked at 15 minutes and was associated with depolymerized tubulin. Staining: Panel A shows STED data (AKT-pS473, red channel) collected simultaneously with confocal signal (a-tubulin, green channel). Upon stimulation of cells with EGF, a rapid activation of AKT is observed (Panel B) along with a coincident change in the tubulin organization (yellow signal), as well as an extensive cell shape-change (cell membrane folding) and accumulation of AKTpS473 at the cell periphery.
High resolution STED immunofluorescence nanoscopy of Mouse anti-AKT pS473 antibody. Tissue: A431 cells. The merge images (A) and at high magnification (B) show phosphorylated AKT colocalized with the distal microtubules. Fixation: 4% paraformaldehyde for 5 min and after washes blocked with 10% NGS/0.2% Triton X-100 for 30 min. Antigen retrieval: serum deprivation for 12 h. Primary antibody: AKT pS473 antibody at 10 ug/mL and A Alpha-tubulin (cyan) (p/n 600-401-880) at 1.4 ug/mL for 1 h at RT. Secondary antibody: Atto 647N anti-Mouse IgG (ATTO TEC GmbH), and DyLight™488 anti-Rabbit IgG (p/n 611-141-122) were used at 1.0 ug/mL for 1h at RT for indirect detection. Localization: AKT pS473 is in the cytoplasm and also organized at the periphery of the cell. Staining: AKT pS473 as red signal with bis-benzimide (blue) nuclear counterstain.