AKT1/2/3 | HUABIO HA750179 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

AKT1/2/3

catalog :

HA750179

quantity :

100μl

price :

649.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

ST48-09

reactivity :

human, mouse, rat

application :

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of AKT1/2/3 on different lysates with Rabbit anti-AKT1/2/3 antibody (HA750179) at 1/5,000 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) Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 24 seconds; 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 (HA750179) at 1/5,000 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.

image 2 :

AKT1/2/3 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA750179 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750179 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: HA750179 IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of HA750179 in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 23 seconds; ECL: K1801

image 3 :

Immunocytochemistry analysis of MCF7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

product information

SKU :

HA750179

Target name :

AKT1/2/3

Species reactivity :

Human,Mouse,Rat,Monkey

Applications :

WB,IF-Cell,IF-Tissue,IHC-P,IP,FC,IHC-Fr

Conjugate :

Non-conjugated

Immunogen :

Recombinant protein within human AKT3 aa 300-479.

Uniprot id :

P31749>SwissProt: P31749 Human;SwissProt: P31751 Human;SwissProt: Q9Y243 Human;SwissProt: P31750 Mouse;SwissProt: Q60823 Mouse;SwissProt: Q9WUA6 Mouse;SwissProt: P47196 Rat;SwissProt: P47197 Rat;SwissProt: Q63484 Rat

Host :

Rabbit

Clone number :

ST48-09

Isotype :

IgG

Size :

100μl

List Price :

649.00 USD

Storage Buffer :

PBS (pH7.4).

Form :

Liquid

Storage Instruction :

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

MCF7 cell lysate, U-2 OS cell lysate, Jurkat cell lysate, C6 cell lysate, mouse heart tissue lysate, mouse testis tissue lysate, rat heart tissue lysate, rat testis tissue lysate, MCF7, RAW264.7, C6, human brain tissue, human lung tissue, mouse brain tissue, mouse lung tissue, rat brain tissue, rat lung tissue, mouse hippocampus tissue, mouse cerebral cortex tissue.

Molecular wt :

Predicted band size: 56 kDa

Subcellular location :

Cell membrane, Cytoplasm, Membrane, Nucleus.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:5,000-1:10,000 ;IF-Cell: 1:100-1:200 ;IF-Tissue: 1:500-1:1,000 ;IHC-P: 1:2,000 ;FC: 1:1,000 ;IP: 1-2μg/sample ;IHC-Fr: 1:100

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA750179_4.jpg

Pic legend4 :

Immunocytochemistry analysis of RAW264.7 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/HA750179_5.jpg

Pic legend5 :

Immunocytochemistry analysis of C6 cells labeling AKT1/2/3 with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/HA750179_6.jpg

Pic legend6 :

Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/HA750179_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/HA750179_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/HA750179_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/HA750179_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img11 :

https://storage.huabio.cn/huabio/productImg/HA750179_11.jpg

Pic legend11 :

Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-AKT1/2/3 antibody (HA750179) 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 (HA750179) 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.

Pic img12 :

https://storage.huabio.cn/huabio/productImg/HA750179_12.jpg

Pic legend12 :

Application: IHC-Fr 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.

Pic img13 :

https://storage.huabio.cn/huabio/productImg/HA750179_13.jpg

Pic legend13 :

Application: IHC-Fr 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.

Pic img14 :

https://storage.huabio.cn/huabio/productImg/HA750179_14.jpg

Pic legend14 :

Flow cytometric analysis of MCF7 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750179, 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).

Pic img15 :

https://storage.huabio.cn/huabio/productImg/HA750179_15.jpg

Pic legend15 :

Flow cytometric analysis of RAW264.7 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750179, 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).

Pic img16 :

https://storage.huabio.cn/huabio/productImg/HA750179_16.jpg

Pic legend16 :

Flow cytometric analysis of C6 cells labeling AKT1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750179, 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).

more info or order :

HUABIO product webpage