AKT1/2/3 Recombinant Rabbit Monoclonal Antibody [JE75-09]
cat.: HA721870
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat, Monkey |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | JE75-09 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 56 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human AKT1/2/3 aa 231-480 / 480. |
| Positive control: | HeLa cell lysate, MCF7 cell lysate, A549 cell lysate, U-2 OS cell lysate, NIH/3T3 cell lysate, MCF7, COS-1 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Mouse testis tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, Rat testis tissue lysate, RAW264.7, C6. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:2,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P31749 Human | P31751 Human | Q9Y243 Human | P31750 Mouse | Q60823 Mouse | Q9WUA6 Mouse | P47196 Rat | P47197 Rat | Q63484 Rat |
| Alternative names: | AKT AKT1 AKT1 kinase AKT1m AKT2 AKT2 kinase Akt3 AKT3_HUMAN CAKT CWS6 DKFZp434N0250 HIHGHH kinase Akt1 MGC99656 MPPH Murine thymoma viral (v-akt) homolog 2 pan-akt PKB ALPHA PKB PKB beta PKB gamma PKB-GAMMA PKB/Akt PKBALPHA PKBB PKBBETA PKBG PKBGAMMA PRKBA PRKBB PRKBG Protein kinase Akt 2 Protein kinase Akt-2 Protein kinase Akt-3 Protein kinase B alpha Protein kinase B Protein kinase B beta Protein kinase B gamma Proto oncogene c Akt Proto-oncogene c-Akt RAC ALPHA RAC alpha serine/threonine protein kinase RAC RAC BETA RAC beta serine/threonine protein kinase RAC PK alpha RAC PK beta rac protein kinase alpha rac protein kinase beta RAC-ALPHA RAC-alpha serine/threonine-protein kinase RAC-beta serine/threonine-protein kinase RAC-gamma RAC-gamma serine/threonine-protein kinase RAC-PK-alpha RAC-PK-beta RAC-PK-gamma RACALPHA RACalpha serine/threonine kinase RACBETA RACgamma...... |
Images
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Fig1:
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody (HA721870) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: A549 cell lysate Lane 4: U-2 OS cell lysate Lane 5: NIH/3T3 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 30 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 (HA721870) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of MCF7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA721870) 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 (HA721870) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody (HA721870) at 1/2,000 dilution. Lane 1: MCF7 cell lysate Lane 2: A549 cell lysate Lane 3: U-2 OS cell lysate Lane 4: COS-1 cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: RAW264.7 cell lysate Lane 7: C6 cell lysate Lane 8: PC-12 cell lysate Lane 9: Mouse brain tissue lysate Lane 10: Mouse heart tissue lysate Lane 11: Mouse testis tissue lysate Lane 12: Rat brain tissue lysate Lane 13: Rat heart tissue lysate Lane 14: Rat testis tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 2 minutes; ECL: K1801; 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 (HA721870) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig4:
AKT1/2/3 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA721870 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA721870 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: HA721870 IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of HA721870 in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 6 seconds; ECL: K1801 |
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Fig5:
Immunocytochemistry analysis of RAW264.7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA721870) 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 (HA721870) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig6:
Immunocytochemistry analysis of C6 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA721870) 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 (HA721870) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig7:
Flow cytometric analysis of C6 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721870, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig8:
Flow cytometric analysis of RAW264.7 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721870, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig9:
Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody (HA721870) at 1/2,000 dilution. Lane 1: MCF7-si NT cell lysate Lane 2: MCF7-si AKT1/2/3 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 7 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 (HA721870) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.