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Anti-AKT1 Antibody (phospho-Ser473, clone 17F6.B11) LS-C153997

Ordering

Wt. Vol. Conc. Price
100 µg - 1.02 mg/ml $405
Inquire for larger quantities

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Most Popular AKT1 Antibodies

Anti-AKT1 Antibody (phospho-Ser473) IHC-plus™ LS-B1183
Rabbit Polyclonal to Human AKT1
Human, Mouse, Rat
IHC - Paraffin, ICC, Immunofluorescence, Western blot, ELISA
Unconjugated
Immunohistochemistry Image
Anti-AKT1 Antibody (aa121-170) IHC-plus™ LS-B1510
Rabbit Polyclonal (IgG) to Human AKT1
Human, Monkey, Mouse, Rat, Bovine, Dog, Goat, Guinea pig, Hamster, Horse, Sheep, Chicken, Xenopus
IHC - Paraffin, Western blot
Unconjugated
Immunohistochemistry Image
Anti-AKT1 Antibody (phospho-Thr450, PE) LS-C252845
Rabbit Polyclonal (IgG) to Human AKT1
Human
ELISA, Dot Blot
PE Conjugated

100% Guaranteed 100% Guaranteed
Mouse Monoclonal [clone 17F6.B11] (IgG1) to Human AKT1
Human, Monkey, Mouse, Rat
IHC - Paraffin, Immunofluorescence, Western blot, ELISA
Unconjugated

Details

Human AKT1
Mouse
Human, Monkey, Mouse, Rat (tested or 100% immunogen sequence identity)
IgG1 Monoclonal [17F6.B11]
Unconjugated
Protein A purified
Unmodified

Applications

  • IHC - Paraffin (20 µg/ml)
  • Immunofluorescence (1:500 - 1:3000)
  • Western blot (1:500 - 1:3000)
  • ELISA

Specificity and Use

AKT1 antibody was raised against anti-AKT pS473 (MOUSE) Monoclonal Antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein.
pSer473
This antibody is specific for human and mouse AKT protein phosphorylated 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. Cross-reactivity with AKT from other sources has not been determined. Cross-reactivity with AKT2 and AKT3 has not been determined.
Expect a band approximately 56 kD in size corresponding to phosphorylated AKT protein by western blotting in the appropriate cell lysate or extract. This phospho-specific monoclonal antibody reacts with human and mouse AKT pS473 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. Specific conditions for reactivity should be optimized by the end user. For immunohistochemistry use formalin-fixed paraffin-embedded sections. No pre-treatment of sample is required.

Packaging

0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 0.01% sodium azide, sterile filtered
Short term 4°C, long term aliquot and store at -20°C, avoid freeze thaw cycles.
For research use only.

About AKT1

P31749 NM_005163 NP_005154.2

AKT1 Antibody, AKT Antibody, AKT-1 Antibody, C-AKT Antibody, Pan-AKT Antibody, PKB alpha Antibody, PKBalpha Antibody, Proto-oncogene c-Akt Antibody, Protein kinase B Antibody, Rac protein kinase alpha Antibody, PRKBA Antibody, Protein kinase B alpha Antibody, RAC-PK-alpha Antibody, PKB Antibody, PKB-ALPHA Antibody, RAC Antibody, RAC-ALPHA Antibody

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.

Immunohistochemistry

Immunohistochemistry
AKT pS473 Monoclonal Antibody - Immunohistochemistry. anti-AKT pS473 monoclonal antibody in immunohistochemistry shows detection of phosphorylated AKT pS473 in human prostate tissue. The antibody was used at 20 ug/mL. The staining is much stronger than the weak basal level of phosphorylation in normal prostate tissue. Tissue was formalin fixed and paraffin embedded. No pre-treatment of sample was required.

Immunofluorescence

Immunofluorescence
AKT pS473 Antibody - Immunofluorescence Microscopy. Anti-AKT pS473 antibody used in confocal microscopy shows detection of changes in AKT pS473 localization in EGF treated A431 cells. A Leica TCS SP5 was used to detect tubulin (cyan) stained with DyLight 488 Goat anti-Rabbit IgG, and AKT (red) stained with MAb anti-AKT pS473 and detected with atto-647N anti-Mouse IgG (Active Motif). The images show a weak diffuse staining of AKT in serum starved resting cells (No treatment), and a marked activation and migration of AKT to the periphery of the cells upon stimulation with the mitogen EGF (+ EGF 15 min).

Immunofluorescence

Immunofluorescence
AKT pS473 Monoclonal Antibody - Immunofluorescence Microscopy. High resolution STED immunofluorescence nanoscopy showing detection of phosphorylated AKT. anti-AKT pS473 monoclonal antibody LS-C153997 was used at 10 ug/mL for 1 h at RT to stain A431 cells after serum deprivation for 12 h. Cells were fixed in 4% paraformaldehyde for 5 min and after washes blocked with 10% NGS/0.2% Triton X-100 for 30 min. Phosphorylated AKT (red) is observed to be localized in the cytoplasm and also organized at the periphery of the cell. Sequential staining at 1.4 ug/mL for 1 h at RT with rabbit anti-a tubulin (cyan) (LS-B722) is also shown. The merge images (A) and at high magnification (B) show phosphorylated AKT colocalized with the distal microtubules. High magnification images (D, E and F) show phosphorylated AKT appears to be colocalized to microtubules. Images were taken with a 100x objective on a Leica TCS STED (Stimulated Emission and Depletion), Leica Microsystems, Inc. Exton, PA, USA. Atto 647N anti-Mouse IgG (ATTO TEC GmbH), and DyLight488 anti-Rabbit IgG (p/n were used at 1.0 ug/mL for 1h at RT for indirect detection. Prolong antifade reagent (Invitrogen) was used to mount slides. Personal Communication, Myriam Gastard, Leica Microsystems Inc, Exton, PA (USA).

Western blot

Western blot
AKT pS473 Monoclonal Antibody - Western Blot. Anti-AKT pS473 by western blot shows detection of phosphorylated AKT (indicated by arrowhead at ~56 kD) on PDGF stimulated NIH/3T3 cell lysates (lane 2). No reactivity is seen for non-phosphorylated AKT in untreated cells (lane 1). Each lane contained approximately 10 ug of lysate. All samples were loaded on to a 4-20% gradient gel for separation. After electrophoresis, the gel was blocked with 5% BLOTTO (B501-0500 in TBS for 90 min at RT. The membrane was probed with the primary antibody at a 1:10000 dilution in TBS with 0.05% Tween-20 with 1% BSA, for 1 h at 4° C. For detection HRP conjugated Gt-a-Mouse IgG (p/n LS-C60680) was used at a 1:20000 dilution for 1 h at 4° C with FemtoMax enhanced chemiluminescent reagent (p/n FEMTOMAX-100). Images were captured using 2X2 binning for 10-20 sec using a BioSpectrum Imaging System (UVP Ltd.).

Western blot

Western blot
AKT pS473 Monoclonal Antibody - Western Blot. Anti-AKTpS473 Antibody in western blot showing detection of AKTpS473 in A431 cells. Epidermal growth factor (EGF) was used to stimulate EGF receptor (EGFR) in A431 cells. Cells were stimulated for 15 min with EGF. The Western blot was blocked with Blocking Buffer for Fluorescent Western Blot p/n MB-070, incubated with DyLight649 Conjugated Anti-AKT pS473 Monoclonal Antibody p/n and detected using the BioRad VersaDoc MP4000 system.

Western blot

Western blot
AKT pS473 Monoclonal Antibody - Western Blot. Anti-AKT pS473 Monoclonal Antibody - Western Blot#Anti-Akt pS473 antibody by fluorescent western blot shows simultaneous detection of unphosphorylated and phosphorylated Akt1 present in serum starved and PDGF stimulated NIH/3T3 whole cell lysates. Lane 1, unstimulated NIH/3T3 lysates contain inactive unphosphorylated Akt1, green band. Lane 2, PDGF stimulated NIH/3T3 lysate contains both inactive (green band) and activated phosphorylated Akt1 (red band). Both lanes were probed with rabbit anti-Akt (pan) and mouse anti-Akt pS473 specific antibodies. This was followed by detection with DyLight 549 conjugated anti-rabbit IgG (green) and DyLight 649 conjugated anti-mouse IgG (red) secondary antibodies.

Requested From: United States
Date Requested: 12/9/2016

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