WB
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. Lane 1: MCF7 cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: U-2 OS cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: NIH/3T3 cell lysate (20 µg/Lane) Lane 6: RAW264.7 cell lysate (20 µg/Lane) Lane 7: C6 cell lysate (20 µg/Lane) Lane 8: PC-12 cell lysate (20 µg/Lane) Lane 9: Mouse brain tissue lysate (20 µg/Lane) Lane 10: Mouse heart tissue lysate (20 µg/Lane) Lane 11: Mouse testis tissue lysate (20 µg/Lane) Lane 12: Rat brain tissue lysate (20 µg/Lane) Lane 13: Rat heart tissue lysate (20 µg/Lane) Lane 14: Rat testis tissue lysate (20 µg/Lane) Exposure time: 24 seconds; 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.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 (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of MCF7 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 iFluor™ 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 Rabbit anti-AKT1/2/3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-AKT1/2/3 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: Rabbit anti-AKT1/2/3 antibody IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-AKT1/2/3 antibody in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 23 seconds.IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-AKT1/2/3 antibody= at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.IF-F
Species: Mouse, Site: Hippocampus, Sample: Frozen section, Antibody concentration: 1/100, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.IF-F
Species: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/100, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.| Product Name | AKT1/2/3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human AKT3 aa 300-479. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-FIHCIPWB |
| WB:1:2000-1:5000 IHC:1:2000 ICC/IF:1:50-1:200 IF-F:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | AKT1/2/3 |
|---|---|
| Gene Synonyms | AKT PKB PKB-ALPHA PRKBA RAC RAC-ALPHA HIHGHH PKBB PKBBETA PRKBB RAC-BETA MPPH MPPH2 PKB-GAMMA PKBG PRKBG RAC-PK-gamma RAC-gamma STK-2 |
| Gene Full Name | AKT serine/threonine kinase 3 |
| Gene Summary | The protein encoded by this gene is a member of the AKT, also called PKB, serine/threonine protein kinase family. AKT kinases are known to be regulators of cell signaling in response to insulin and growth factors. They are involved in a wide variety of biological processes including cell proliferation, differentiation, apoptosis, tumorigenesis, as well as glycogen synthesis and glucose uptake. This kinase has been shown to be stimulated by platelet-derived growth factor (PDGF), insulin, and insulin-like growth factor 1 (IGF1). Alternatively splice transcript variants encoding distinct isoforms have been described. [provided by RefSeq, Jul 2008] |
| 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 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 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-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 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-gamma antibody RAC-gamma serine/threonine-protein kinase 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 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 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) | 56kDa |
| Cellular Localization | Cell membrane, Cytoplasm, Membrane, Nucleus. |
WB
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. Lane 1: MCF7 cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: U-2 OS cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: NIH/3T3 cell lysate (20 µg/Lane) Lane 6: RAW264.7 cell lysate (20 µg/Lane) Lane 7: C6 cell lysate (20 µg/Lane) Lane 8: PC-12 cell lysate (20 µg/Lane) Lane 9: Mouse brain tissue lysate (20 µg/Lane) Lane 10: Mouse heart tissue lysate (20 µg/Lane) Lane 11: Mouse testis tissue lysate (20 µg/Lane) Lane 12: Rat brain tissue lysate (20 µg/Lane) Lane 13: Rat heart tissue lysate (20 µg/Lane) Lane 14: Rat testis tissue lysate (20 µg/Lane) Exposure time: 24 seconds; 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
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 (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of MCF7 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 iFluor™ 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 Rabbit anti-AKT1/2/3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-AKT1/2/3 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: Rabbit anti-AKT1/2/3 antibody IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-AKT1/2/3 antibody in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 23 seconds.
IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-AKT1/2/3 antibody= at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-AKT1/2/3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
IF-F
Species: Mouse, Site: Hippocampus, Sample: Frozen section, Antibody concentration: 1/100, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.
IF-F
Species: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/100, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.| Application Notes | WB:1:2000-1:5000 IHC:1:2000 ICC/IF:1:50-1:200 IF-F:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
