Phospho-AKT1(Ser473) 抗体 (YA227)

Phospho-Akt1(Ser473) Antibody is a non-conjugated and Rabbit origined monoclonal antibody about 56 kDa, targeting to Phospho-Akt1(Ser473). It can be used for WB,ICC/IF,IHC-P assays with tag free, in the background of Human, Mouse, Rat, Dog.

Free

P31749

Predicted 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: 1:5,000-1:10,000 ;IF-Cell: 1:100-1:1,000 ;IHC-P: 1:200-1:1,000; IF-Tissue: 1:200-1:500; IHC-Fr: 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.