AKT Monoclonal antibody, PBS Only (Detector)

• Featured Product
• KD/KO Validated

AKT Monoclonal Antibody for WB, IP, IF, FC, IHC, Indirect ELISA, Cytometric bead array

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IP, IF, FC, IHC, Indirect ELISA, Cytometric bead array

Conjugate

Unconjugated

CloneNo.

2C5D1

Cat no : 60203-2-PBS

Synonyms

AKT, AKT (pan), AKT pan, AKT1, AKT1/2/3, pan AKT, PKB, PKB ALPHA, PRKBA, Protein kinase B, Proto oncogene c Akt, RAC, RAC ALPHA, RAC PK alpha, total AKT

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Validation Data Gallery

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  WB analysis of A549 using 60203-2-Ig (same clone as 60203-2-PBS)

  WB result of AKT antibody (60203-2-Ig; 1:5000; incubated at room temperature for 1.5 hours) with sh-Control and sh-AKT transfected A549 cells. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis using 60203-2-Ig (same clone as 60203-2-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of HEK-293 using 60203-2-Ig (same clone as 60203-2-PBS)

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) and 66444-1-Ig(AKT1-phospho-S473 Antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of rat liver using 60203-2-Ig (same clone as 60203-2-PBS)

  rat liver tissue were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of mouse brain using 60203-2-Ig (same clone as 60203-2-PBS)

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of ROS1728 using 60203-2-Ig (same clone as 60203-2-PBS)

  ROS1728 cells were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of HEK-293 using 60203-2-Ig (same clone as 60203-2-PBS)

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of NIH/3T3 using 60203-2-Ig (same clone as 60203-2-PBS)

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  WB analysis of RAW 264.7 using 60203-2-Ig (same clone as 60203-2-PBS)

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IP experiment of mouse brain using 60203-2-Ig (same clone as 60203-2-PBS)

  IP result of anti-AKT (IP:60203-2-Ig, 5ug; Detection:60203-2-Ig 1:1000) with mouse brain tissue lysate 4000ug. This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60203-2-Ig (AKT Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 60203-2-Ig (AKT Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human cervical cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human cervical cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 10x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human cervical cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human cervical cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 40x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 10x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 40x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 10x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 60203-2-Ig(AKT antibody) at dilution of 1:50 (under 40x lens). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  IF Staining of MCF-7 using 60203-2-Ig (same clone as 60203-2-PBS)

  Immunofluorescent analysis of (-20°C Methanol) fixed MCF-7 cells using AKT antibody (60203-2-Ig, Clone: 2C5D1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

• 

  FC experiment of Jurkat using 60203-2-Ig (same clone as 60203-2-PBS)

  1X10^6 Jurkat cells were intracellularly stained with 0.2 ug Anti-Human AKT (60203-2-Ig, Clone:2C5D1) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Mouse IgG1 Isotype Control (66360-1-Ig, Clone: T1F8D3F10) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C). This data was developed using the same antibody clone with 60203-2-PBS in a different storage buffer formulation.

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  Cytometric bead array in cell lysate using MP50177-1

  Cytometric bead array in cell lysate using MP50177-1, Phospho-AKT(Ser473) Monoclonal Matched Antibody Pair, PBS Only. Capture antibody: 66444-1-PBS. Detection antibody: 60203-2-PBS. Cell lysate: Non-treated HEK293 and Calyculin A treated HEK293(30μg/well). Non-related target OAT Monoclonal Matched Antibody Pair (MP50109-1P) was served as control.

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Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

60203-2-PBS targets AKT as part of a matched antibody pair:

MP50177-1: 66444-1-PBS capture and 60203-2-PBS detection (validated in Cytometric bead array)

Unconjugated mouse monoclonal antibody pair in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: AKT fusion protein Ag16695
• Full Name: v-akt murine thymoma viral oncogene homolog 1
• Calculated Molecular Weight: 56 kDa
• Observed Molecular Weight: 56-62 kDa
• GenBank Accession Number: BC000479
• Gene Symbol: AKT1
• Gene ID (NCBI): 207
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS Only
• Storage Conditions: Store at -80°C.

Background Information

The serine-threonine protein kinase AKT1 is catalytically inactive in serum-starved primary and immortalized fibroblasts. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. This antibody detects all the members of AKT with/without phospho- modification.