WB
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 4-20% 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 at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of C6 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of C6 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).FC
Flow cytometric analysis of RAW264.7 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
AKT1/2/3 was immunoprecipitated from 0.2 mg MCF7 cell lysate with AKT1/2/3 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using AKT1/2/3 Recombinant Rabbit Monoclonal Antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: AKT1/2/3 Recombinant Rabbit Monoclonal Antibody IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of AKT1/2/3 Recombinant Rabbit Monoclonal Antibody in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 6 seconds.| Product Name | AKT1/2/3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human AKT1/2/3 aa 231-480 / 480. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:2000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | AKT1/AKT2/AKT3 |
|---|---|
| Gene Synonyms | AKT PKB RAC PRKBA PKB-ALPHA RAC-ALPHA |
| Gene Full Name | AKT serine/threonine kinase 1 |
| Gene Summary | This gene encodes one of the three members of the human AKT serine-threonine protein kinase family which are often referred to as protein kinase B alpha, beta, and gamma. These highly similar AKT proteins all have an N-terminal pleckstrin homology domain, a serine/threonine-specific kinase domain and a C-terminal regulatory domain. These proteins are phosphorylated by phosphoinositide 3-kinase (PI3K). AKT/PI3K forms a key component of many signalling pathways that involve the binding of membrane-bound ligands such as receptor tyrosine kinases, G-protein coupled receptors, and integrin-linked kinase. These AKT proteins therefore regulate a wide variety of cellular functions including cell proliferation, survival, metabolism, and angiogenesis in both normal and malignant cells. AKT proteins are recruited to the cell membrane by phosphatidylinositol 3,4,5-trisphosphate (PIP3) after phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) by PI3K. Subsequent phosphorylation of both threonine residue 308 and serine residue 473 is required for full activation of the AKT1 protein encoded by this gene. Phosphorylation of additional residues also occurs, for example, in response to insulin growth factor-1 and epidermal growth factor. Protein phosphatases act as negative regulators of AKT proteins by dephosphorylating AKT or PIP3. The PI3K/AKT signalling pathway is crucial for tumor cell survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating AKT1 which then phosphorylates and inactivates components of the apoptotic machinery. AKT proteins also participate in the mammalian target of rapamycin (mTOR) signalling pathway which controls the assembly of the eukaryotic translation initiation factor 4F (eIF4E) complex and this pathway, in addition to responding to extracellular signals from growth factors and cytokines, is disregulated in many cancers. Mutations in this gene are associated with multiple types of cancer and excessive tissue growth including Proteus syndrome and Cowden syndrome 6, and breast, colorectal, and ovarian cancers. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jul 2020] |
| Alternative Names | AKT antibody AKT1 antibody AKT1 kinase antibody AKT1m antibody AKT2 antibody AKT2 kinase antibody Akt3 antibody AKT3_HUMAN antibody CAKT antibody CWS6 antibody DKFZp434N0250 antibody HIHGHH antibody kinase Akt1 antibody MGC99656 antibody MPPH antibody Murine thymoma viral (v-akt) homolog 2 antibody pan-akt antibody PKB ALPHA antibody PKB antibody PKB beta antibody PKB gamma antibody PKB-GAMMA antibody PKB/Akt antibody PKBALPHA antibody PKBB antibody PKBBETA antibody PKBG antibody PKBGAMMA antibody PRKBA antibody PRKBB antibody PRKBG antibody Protein kinase Akt 2 antibody Protein kinase Akt-2 antibody Protein kinase Akt-3 antibody Protein kinase B alpha antibody Protein kinase B antibody Protein kinase B beta antibody Protein kinase B gamma antibody Proto oncogene c Akt antibody Proto-oncogene c-Akt antibody RAC ALPHA antibody RAC alpha serine/threonine protein kinase antibody RAC antibody RAC BETA antibody RAC beta serine/threonine protein kinase antibody RAC PK alpha antibody RAC PK beta antibody rac protein kinase alpha antibody rac protein kinase beta antibody RAC-ALPHA antibody RAC-alpha serine/threonine-protein kinase antibody RAC-beta serine/threonine-protein kinase antibody RAC-gamma antibody RAC-gamma serine/threonine-protein kinase antibody RAC-PK-alpha antibody RAC-PK-beta antibody RAC-PK-gamma antibody RACALPHA antibody RACalpha serine/threonine kinase antibody RACBETA antibody RACgamma antibody RACgamma serine/threonine protein kinase antibody RACPKgamma antibody serine threonine protein kinase antibody STK 2 antibody STK-2 antibody STK2 antibody thymoma viral proto oncogene 1 antibody thymoma viral proto oncogene antibody V akt murine thymoma viral oncogene homolog 1 antibody V akt murine thymoma viral oncogene homolog 2 antibody V akt murine thymoma viral oncogene homolog 3 (protein kinase B gamma) antibody V akt murine thymoma viral oncogene homolog 3 antibody V-AKT murine thymoma viral oncogene homolog 1 antibody V-AKT murine thymoma viral oncogene homolog 2 antibody V-AKT murine thymoma viral oncogene homolog 3 antibody vakt murine thymoma viral oncogene homolog 1 antibody vakt murine thymoma viral oncogene homolog 2 antibody vakt murine thymoma viral oncogene homolog 3 antibody |
| Molecular Weight(MW) | 56 kDa |
| Cellular Localization | Cytoplasm, Nucleus, Cell membrane. |
WB
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 4-20% 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 at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-AKT1/2/3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of C6 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
FC
Flow cytometric analysis of RAW264.7 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
AKT1/2/3 was immunoprecipitated from 0.2 mg MCF7 cell lysate with AKT1/2/3 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using AKT1/2/3 Recombinant Rabbit Monoclonal Antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: AKT1/2/3 Recombinant Rabbit Monoclonal Antibody IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of AKT1/2/3 Recombinant Rabbit Monoclonal Antibody in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 6 seconds.| Application Notes | WB:1:2000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
