antibody

Anti-Phospho-Akt(Ser473) Antibody [SY28-05]

ET1607-73TR

20 uL

99 USD

monoclonal

domestic rabbit

nonconjugated

SY28-05

human, mouse, rat

western blot, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

Anti-Phospho-Akt(Ser473) Antibody [SY28-05]

Human,Mouse,Rat

WB,IF-Cell,IF-Tissue,IHC-P,IHC-Fr

Non-conjugated

Synthetic phospho-peptide corresponding to residues surrounding Ser473 of human Akt1.

SwissProt: P31749 Human;SwissProt: P31751 Human;SwissProt: Q9Y243 Human;SwissProt: P31750 Mouse

Rabbit

SY28-05

IgG

20 uL

99 USD

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

Liquid

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Protein A affinity purified.

Recombinant Rabbit monoclonal Antibody

MCF7 treated with 100nM Calyculin A for 30 minutes whole cell lysate, NIH/3T3 treated with 100ng/mL PDGF for 1 hour whole cell lysate, C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate, mouse lung tissue, hela cell lysates, NIH/3T3 treated with 100nM Calyculin A for 30minutes whole cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30minutes and 10ng/mL hPDGF for 10minutes whole cell lysate, NIH/3T3.

Predicted band size: 56 kDa

Cytoplasm, Nucleus, Cell membrane.

1 mg/mL.

WB: 1:5,000 ;IF-Cell: 1:50-1:200 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:50-1:200 ;IHC-Fr: 1:100

ET1607-73_2.jpg

Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Phospho-Akt(Ser473) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-73, 1/50) for 30 minutes at room temperature. 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.

ET1607-73_3.jpg

Western blot analysis of Phospho-Akt(Ser473) on Hela cell lysates. Lane 1: Hela cells were starved overnight, whole cell lysates, 10 μg/lane. Lane 2: Hela cells were starved overnight, then treated with 150 nM insulin for 15 mins, whole cell lysates, 10 μg/lane. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Phospho-Akt(Ser473) (ET1607-73, 1/500) , Anti-Akt1 antibody ( ET1609-47, 1/500) and Anti-GAPDH antibody (ET1601-4, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 1 minute 28 seconds

ET1607-73_4.jpg

Western blot analysis of Phospho-Akt(Ser473) on different lysates with Rabbit anti-Phospho-Akt(Ser473) antibody (ET1607-73) at 1/500 dilution. Lane 1: NIH/3T3 whole cell lysate Lane 2: NIH/3T3 treated with 100nM Calyculin A for 30minutes whole cell lysate Lane 3: NIH/3T3 treated with 100nM Calyculin A for 30minutes and 10ng/mL hPDGF for 10minutes whole cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 2 minutes; 10% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-73) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

ET1607-73_5.jpg

ICC staining of Phospho-Akt(Ser473) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-73, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1607-73_6.jpg

Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Phospho-Akt(Ser473) with Rabbit anti-Phospho-Akt(Ser473) antibody (ET1607-73). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1607-73, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

ET1607-73_7.jpg

Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Phospho-Akt(Ser473) with Rabbit anti-Phospho-Akt(Ser473) antibody (ET1607-73). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1607-73, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.