Phospho-AKT1(Ser473) 抗体 (YA227)

Phospho-AKT1(Ser473) Antibody (YA227) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-AKT1(Ser473).

AKT1 蛋白与 AKT2 和 AKT3 一样是 AKT 激酶家族的成员，在调节代谢、增殖、细胞存活、生长和血管生成等多种细胞过程中发挥着关键作用。AKT1 通过众多下游底物的丝氨酸和/或苏氨酸磷酸化发挥作用，参与 100 多种报道的候选底物的调节，这证明了其多方面的性质。值得注意的是，它协调胰岛素诱导的 SLC2A4/GLUT4 葡萄糖转运蛋白易位，影响葡萄糖摄取。此外，AKT1 通过磷酸化 GSK3A 和 GSK3B 来控制葡萄糖以糖原形式储存，抑制其激酶活性。它参与多种途径，包括通过 mTORC1 信号途径介导胰岛素刺激的蛋白质合成以及调节 FOXO 因子，影响细胞定位。AKT1 还影响 NF-kappa-B 依赖性基因转录并正向调节 CREB1 活性，有助于促生存基因的转录。此外，AKT1 通过磷酸化 ATP 柠檬酸裂解酶 (ACLY) 并激活环核苷酸磷酸二酯酶 (PDE3B) 的 3B 同工型，从而在脂质代谢中发挥作用，从而降低环 AMP 水平并抑制脂肪分解。除了这些功能之外，AKT1 还参与细胞迁移、粘附组装和分解、胎盘发育以及对生长因子的多种细胞反应，包括血小板源性生长因子 (PDGF)、表皮生长因子 (EGF)、胰岛素和胰岛素样因子生长因子 I (IGF-I)。最近发现 AKT1 在 SPATA13 介导的细胞迁移和粘附调节中的作用增加了其功能的复杂性，突出了 AKT1 在细胞稳态和对环境信号的响应中的核心作用。

Free

P31749

Predicted band size: 56 kDa; Observed band size: 56 kDa

Protein A affinity purified.

Cytoplasm, Nucleus, Cell membrane.

Non-conjugated

Phosphorylated

AB_3102397

Neuroscience

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat, Dog

WB: 1:5000-1:10000; ICC: 1:100-1:1000; IHC-P: 1:200-1:1000; IF-Tissue: 1:200-1:500; IHC-F: 1:100

• 
  Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7(lane 3(20μg) and Jurkat(lane 4(20ug) using Phospho-AKT1(HY-P80276 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of C6 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with A Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P

液体

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic phosphopeptide corresponding to residues surrounding Ser473 of Human Akt1.The exact sequence is proprietary to MCE.