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  • Non-treated and Calyculin A treated cell lysates were subjected to SDS PAGE followed by western blot with 67558-1-Ig (Phospho-GSK3B (Ser9) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

    Non-treated and Calyculin A treated cell lysates were subjected to SDS PAGE followed by western blot with 67558-1-Ig (Phospho-GSK3B (Ser9) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

  • Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80596-1-RR (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) subsequently.

    Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80596-1-RR (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) subsequently.

  • Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80596-1-RR (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) subsequently.

    Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80596-1-RR (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) subsequently.

  • Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80698-1-RR (Phospho-TSC2 (Thr1462) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with mTOR antibody (66888-1-Ig) as the loading control.

    Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80698-1-RR (Phospho-TSC2 (Thr1462) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with mTOR antibody (66888-1-Ig) as the loading control.

  • Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81654-1-RR (Phospho-TSC2 (Ser939) antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with mTOR antibody (66888-1-Ig) as the loading control.

    Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81654-1-RR (Phospho-TSC2 (Ser939) antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with mTOR antibody (66888-1-Ig) as the loading control.

  • Non-treated MCF-7 cells and Cobalt Chloride treated MCF-7 cells were subjected to SDS PAGE followed by western blot with 28757-1-AP (Phospho-FOXO1 (Ser319) antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin antibody as loading control.

    Non-treated MCF-7 cells and Cobalt Chloride treated MCF-7 cells were subjected to SDS PAGE followed by western blot with 28757-1-AP (Phospho-FOXO1 (Ser319) antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin antibody as loading control.

  • Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81871-1-RR (Phospho-CREB1 (Ser133) antibody) at dilution of 1:20000 incubated at room temperature for 1 hours. The membrane was stripped and re-blotted with Beta-tubulin antibody as loading control.

    Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81871-1-RR (Phospho-CREB1 (Ser133) antibody) at dilution of 1:20000 incubated at room temperature for 1 hours. The membrane was stripped and re-blotted with Beta-tubulin antibody as loading control.

  • Non-treated NIH/3T3 and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80346-1-RR (Phospho-Caspase 9 (Ser196) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin antibody as loading control.

    Non-treated NIH/3T3 and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80346-1-RR (Phospho-Caspase 9 (Ser196) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin antibody as loading control.

  • Non-treated NIH/3T3 and PDGF (HZ-1215) treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 28791-1-AP (Phospho-XIAP (Ser87) antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

    Non-treated NIH/3T3 and PDGF (HZ-1215) treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 28791-1-AP (Phospho-XIAP (Ser87) antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

  • Phosphatase inhibitor treated and λ phosphatase treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 28847-1-AP (Phospho-ASK1 (Ser83) antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

    Phosphatase inhibitor treated and λ phosphatase treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 28847-1-AP (Phospho-ASK1 (Ser83) antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

  • Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80455-1-RR (Phospho-AKT (Ser473) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

    Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80455-1-RR (Phospho-AKT (Ser473) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

  • Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 67558-1-Ig (GSK3B-phospho-S9 antibody) at dilution of 1:300 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 67558-1-Ig (GSK3B-phospho-S9 antibody) at dilution of 1:300 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 81871-1-RR (Phospho-CREB1 (Ser133) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

    Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 81871-1-RR (Phospho-CREB1 (Ser133) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescent analysis of (-20°C Ethanol) fixed Calyculin A treated PC-3 cells and non-treated PC-3 cells using Phospho-GSK3B (Ser9) antibody (67558-1-Ig, Clone: 1C9E2 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

    Immunofluorescent analysis of (-20°C Ethanol) fixed Calyculin A treated PC-3 cells and non-treated PC-3 cells using Phospho-GSK3B (Ser9) antibody (67558-1-Ig, Clone: 1C9E2 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

  • Immunofluorescent analysis of (4% PFA) fixed PMA treated HEK-293 cells using Phospho-mTOR (Ser2448) antibody (80596-1-RR, Clone: 3L18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

    Immunofluorescent analysis of (4% PFA) fixed PMA treated HEK-293 cells using Phospho-mTOR (Ser2448) antibody (80596-1-RR, Clone: 3L18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

AKT Substrate Expanded Antibody Kit

Cat No. PK30022

Host

Rabbit/Mouse

Reactivity

Target Dependent

Applications

Target Dependent

Conjugate

Unconjugated

Size: 

-/ -

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テクニカルサポートまたはご購入後1年間の交換／補填対応を承ります。詳細はこちらをご覧ください。

Product Information

The AKT Substrate Expanded Antibody Kit provides a cost-effective tool for studying a broad range of proteins that are phosphorylated by AKT. Perfect for signal transduction researchers starting a new project, screening multiple prospective targets, or those who simply require less volume. 

The AKT Substrate Expanded Kit contains 10 antibodies against downstream phosphorylation targets of AKT.


AntigenCatalog No.Host, clonalityTested ReactivityApplicationsVolume
Phospho-GSK3B
(Ser9)		67558-1-Ig            Mouse monoclonalH      WB, IHC, IF            20 uL
Phospho-mTOR
(Ser2448)		80596-1-RR            Rabbit monoclonal            H, RWB, IF           20 uL
Phospho-TSC2
(Thr1462)		80698-1-RR            Rabbit monoclonalHWB          20 uL
Phospho-TSC2
(Ser939)		81654-1-RR            Rabbit monoclonal            H WB         20 uL
Phospho-FOXO1
(Ser319)		28757-1-AP
           		Rabbit polyclonal            H  WB      20 uL            
Phospho-CREB1
(Ser133)		81871-1-RR
           		Rabbit monoclonal            HWB, IHC           20 uL            
Phospho-Caspase 9
(Ser196)		80346-1-RR
           		Rabbit monoclonal            H, M   WB          20 uL            
Phospho-XIAP
(Ser87)		28791-1-AP
           		Rabbit polyclonal            H, M, R   WB            20 uL            
Phospho-ASK1
(Ser83)		28847-1-AP
           		Rabbit polyclonal            HWB            20 uL            
Phospho-AKT
(Ser473)		80455-1-RR
           		Rabbit monoclonal            H       WB          20 uL            


Also see our 'AKT Substrate Essentials Antibody Kit' on the following page  https://www.ptglab.com/products/AKT-Substrate-Essentials-Antibody-Kit-PK30021.htm。

Storage

Store at -20°C. Stable for one year from the date of receipt. 

Background Information

Upon its activation, AKT phosphorylates a number of different substrates, resulting in the activation or inactivation of downstream cellular functions. The phosphorylation of GSK3B at Ser9, inhibits its activity and subsequent glycogen synthesis. Meanwhile, AKT-mediated phosphorylation of mTOR at Ser2448 results in its activation and formation of the mTORC1 complex. TSC2, an inhibitor of mTORC1 activation, is blocked through phosphorylation at Thr1462 and Ser939 residues. FOXO1, a transcription factor invovled in cell cycle arrest and apoptosis, is inhibited by AKT through phosphorylation at Ser319, which leads to its nuclear export. Pro-survival functions are also promoted through inhibiting ASK1 activity via Ser83 phosphorylation and XIAP through Ser87 phosphorylation. Finally, AKT can help to trigger the caspase cascade by phosphorylating Caspase 9 at Ser196. 

Standard Protocols

Click   here    to view our standard protocols for various applications including WB, IP, IHC, IF, FC, and ELISA.

Cited in Article as

pk30022, AKT Substrate Expanded Antibody Kit, Proteintech, IL, USA

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