ab10559 AKT1 Antibody

Western blotting

Recommended dilution: 1:500-1:2000


Immunofluorescence

Recommended dilution: 1:100-1:500


immunocytochemistry

Recommended dilution: 1:100-1:500

 

最佳稀释倍数与浓度应由实验研究人员确认

产品背景

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 (By similarity). Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling (By similarity). 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 (By similarity). Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven (By similarity). AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis (By similarity). Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis (By similarity). Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI(3)P-5 activity (By similarity). The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation (By similarity). Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I (By similarity). Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development (By similarity). Phosphorylates STK4/MST1 at 'Thr-120' and 'Thr-387' leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3. Phosphorylates STK3/MST2 at 'Thr-117' and 'Thr-384' leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation. Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation. Phosphorylates RAF1 at 'Ser-259' and negatively regulates its activity. Phosphorylation of BAD stimulates its pro-apoptotic activity. Phosphorylates KAT6A at 'Thr-369' and this phosphorylation inhibits the interaction of KAT6A with PML and negatively regulates its acetylation activity towards p53/TP53. Phosphorylates palladin (PALLD), modulating cytoskeletal organization and cell motility. Phosphorylates prohibitin (PHB), playing an important role in cell metabolism and proliferation. Phosphorylates CDKN1A, for which phosphorylation at 'Thr-145' induces its release from CDK2 and cytoplasmic relocalization. These recent findings indicate that the AKT1 isoform has a more specific role in cell motility and proliferation. Phosphorylates CLK2 thereby controlling cell survival to ionizing radiation.

Description

Rabbit polyclonal antibody to AKT1

Applications 
WB, IF, ICC.

Immunogen 
AKT1 Antibody detects endogenous levels of total AKT1.

Reactivity 
Human, Mouse, Rat.
可预测：Pig(100%), Zebrafish(%), Horse(%), Dog(%), Chicken(%), Xenopus(%)

Molecular weight
55kDa; 56kD(Calculated).

Host species 
Rabbit

Ig class 
Immunogen-specific rabbit IgG

Purification 
Antigen affinity purification

Full name 
AKT1

Synonyms 
AKT 1; AKT1; AKT1_HUMAN; MGC99656; PKB; PKB-ALPHA; PRKBA; Protein Kinase B Alpha; Protein kinase B; Proto-oncogene c-Akt; RAC Alpha; RAC; RAC-alpha serine/threonine-protein kinase; RAC-PK-alpha;

Storage
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Swissprot 
P31749

Western blot analysis of extracts from 293 cells, using AKT1 Antibody. The lane on the left was treated with blocking peptide. 显示更多 +

Western blot analysis of extracts from PC12(H2O2 treatment), using AKT1 Antibody. The lane on the left was treated with blocking peptide. 显示更多 +

Figure 7 Effect of JHQ on core target proteins abundant in LX-2 cells after CuSO4 treatment. ∗∗∗P < 0.001 and ∗∗P < 0.01 compared with the model group. 显示更多 +

Figure 3 Lycorine changes the activation level of the Akt signaling pathway via RBM10. (A-C) Lycorine significantly decreased the activation level of the Akt signaling pathway. (D) Lycorine significantly increased the protein expression levels of E-cadherin and significantly reduced the protein expression levels of E-cadherin. (E-G) Despite the presence of lycorine, the inhibition of RBM10 significantly increased the activation level of the Akt signaling pathway. (H) Decreasing RBM10 expression induced a significant increase in N-cadherin protein expression. *P<0.05 and **P<0.01. RBM10, RNA binding motif 10; NC, negative control; p, phosphorylated; siRNA, small interfering RNA. 显示更多 +

Fig. 7 Enriched miR-146b-5p in F-MSCs-Exo inhibits PI3K/AKT/mTOR pathway by targeting TRAF6. (A, B) Western blot analysis was performed to detect the impact of F-MSCs-Exo on TRAF6 and the PI3K/AKT/mTOR pathway in rat chondrocytes. (C, D) The expression of TRAF6 was quantitatively analyzed using immunofluorescence staining and ImageJ software (scale bar = 10 μm). (E, F) Direct visualization of chondrocytes treated with nc-inhibitor and miR-146b-5p-inhibitor was performed using Alcian blue staining and safranin staining. (G, H) Western blot analysis was conducted to examine the expressions of TRAF6 and the PI3K/AKT/mTOR pathway in chondrocytes after treatment with nc-inhibitor and miR-146b-5p-inhibitor. (ns, no significant difference; *p 显示更多 +

Figure 8 PF downregulated the protein expression levels of p-AKT1/AKT1, Bcl-2, EGFR, p-mTOR/mTOR, and p-PI3K/PI3K in the colonic mucosa from the dextran sodium sulfate-induced mice. Data are presented as the mean ± SD. *P 显示更多 +

Fig. 10 Expression of GSK-3β and Akt. (A), (B): Validation of RNA-seq data by qRT-PCR. (C), (D), (E): Validation of protein expression by western blot. (means ± SD, n = 3). ***P  显示更多 +

Fig. 8 The expression of core proteins AKT1, ERK2, and CASPASE-9. (A) Relative protein expression levels of AKT1 (a), ERK2 (b), and CASPASE-9 (c) treated with DBD in the MI rat model. Data are presented as the mean ± SD (n = 3). *P < 0.05 and **P < 0.01 versus the sham group; #P < 0.05 and ##P < 0.01 versus the MI group. (B) Immunofluorescence staining to detect the effect of DBD treatment on core proteins expression (original magnifications: 100 × ). (a–c) The expression of AKT1, ERK2, and CASPASE-9. Data are presented as the mean ± SD (n = 3). **P < 0.01 versus the sham group; ##P < 0.01 versus the MI group. 显示更多 +

Fig. 8 The expression of core proteins AKT1, ERK2, and CASPASE-9. (A) Relative protein expression levels of AKT1 (a), ERK2 (b), and CASPASE-9 (c) treated with DBD in the MI rat model. Data are presented as the mean ± SD (n = 3). *P < 0.05 and **P < 0.01 versus the sham group; #P < 0.05 and ##P < 0.01 versus the MI group. (B) Immunofluorescence staining to detect the effect of DBD treatment on core proteins expression (original magnifications: 100 × ). (a–c) The expression of AKT1, ERK2, and CASPASE-9. Data are presented as the mean ± SD (n = 3). **P < 0.01 versus the sham group; ##P < 0.01 versus the MI group. 显示更多 +

Fig. 5. A Apoptosis was examined by flow cytometry. B Apoptosis was determined by mitochondrial membrane potential assay, scale bar: 50 μm. C qPCR was used to detect mRNA expression of PI3K, AKT, mTOR, BCL-2, BAX, caspase-3, and caspase-6. D, E The protein expression levels of p-PI3K, PI3K, p-AKT, AKT, BAX, BCL-2, caspase-3, and cleaved-caspase-3 were examined using a western blot. The full-length blots are presented in Supplementary Figure. ** P  显示更多 +

Figure 8. HGWD alleviated apoptosis after cerebral I/R. (A) TUNEL staining images of five groups. (Scale bar = 50 μm, 400×). (B) HGWD reduced cell apoptosis through PI3K/AKT signalling pathway. Western blotting was utilized to examine the levels of Bcl-2, Bax, caspase-3, cleaved caspase-3, AKT1, p-AKT1, PI3K, and p-PI3K. (C) Percentages of TUNEL positive cells were utilized to evaluate the differences in the five groups. Integrated density data were quantified (D-J). the results (D-H), respectively, showed the levels of Bax, Bcl-2, the ratio of Bcl-2/Bax, caspase-3, and cleaved caspase-3 in the brains of rats for five groups, with β-actin as the internal control. The results of I and J, respectively, showed the ratios of p-AKT1/AKT1 and p-PI3K/PI3K in the brains of rats for five groups. All data were shown as mean ± SD. (n = 3, *p  显示更多 +