AKT1/2/3 | HUABIO HA721870 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

AKT1/2/3

catalog :

HA721870

quantity :

100μl

price :

385.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

JE75-09

reactivity :

human, mouse, rat

application :

western blot, immunoprecipitation, flow cytometry

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 (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; ECL: K1802; 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.

image 2 :

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.

image 3 :

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.

product information

SKU :

HA721870

Target name :

AKT1/2/3

Species reactivity :

Human,Mouse,Rat,Monkey

Applications :

WB,IF-Cell,FC,IP

Conjugate :

Non-conjugated

Immunogen :

Recombinant protein within Human AKT1/2/3 aa 231-480 / 480.

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 :

JE75-09

Isotype :

IgG

Size :

100μl

List Price :

385.00 USD

Storage Buffer :

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Form :

Liquid

Storage Instruction :

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

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.

Molecular wt :

Predicted band size: 56 kDa

Subcellular location :

Cytoplasm, Nucleus, Cell membrane.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:2,000 ;IF-Cell: 1:100 ;FC: 1:1,000 ;IP: 1-2μg/sample

Advanced Validation :

Knockdown (KD)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA721870_4.jpg

Pic legend4 :

AKT1/2/3 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA721870 at 2 µg/10 µl beads. 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

Pic img5 :

https://storage.huabio.cn/huabio/productImg/HA721870_5.jpg

Pic legend5 :

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.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/HA721870_6.jpg

Pic legend6 :

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.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/HA721870_7.jpg

Pic legend7 :

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).

Pic img8 :

https://storage.huabio.cn/huabio/productImg/HA721870_8.jpg

Pic legend8 :

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).

Pic img9 :

https://storage.huabio.cn/huabio/productImg/HA721870_9.jpg

Pic legend9 :

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; 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 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

more info or order :

HUABIO product webpage