bs-6951R [Primary Antibody]
Rabbit  Anti-AKT1+2+3 Polyclonal Antibody
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Host: Rabbit

Target Protein: AKT1+2+3

IR: Immunogen Range:401-480/480

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 207

Swiss Prot: P31749

Source: KLH conjugated synthetic peptide derived from human AKT1/2/3:401-480/480 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background: AKT, also known as protein kinase B (PKB), is a 57 kDa serine/threonine protein kinase. There are three mammalian isoforms of Akt: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH)domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. Its tight control over cell proliferation and cell viability are manifold; overexpression or inappropriate activation of AKT has been seen in many types of cancer. AKT mediates many of the downstream events of phosphatidylinositol 3 kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI3 kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus.

Size: 50ul

Concentration: 1mg/ml

Applications: WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test)
ICC(1:100-500)
IF(1:100-500)

Cross Reactive Species: Human
Mouse
Rat
Chicken
Dog
Pig
Cow
Rabbit
Sheep
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample:Liver (Mouse) Lysate at 40 ug
Primary: Anti-AKT1+2+3(bs-6951R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 56kD
Observed band size: 56kD
Sample:
Lung (Mouse) Lysate at 40 ug
Placenta (Mouse) Lysate at 40 ug
Primary: Anti-AKT1+2+3 (bs-6951R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Sample:
Lane 1: SiHa (Human) Cell Lysate at 30 ug
Lane 2: Cerebrum (Mouse) Lysate at 40 ug
Primary: Anti-AKT1+2+3 (bs-6951R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 59’61 kD
Observed band size: 59 kD
Sample:
Liver (Mouse) Lysate at 40 ug
Lung (Mouse) Lysate at 40 ug
Primary: Anti- AKT1+2+3 (bs-6951R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Sample:
Heart (Mouse) Lysate at 40 ug
Primary: Anti- AKT1+2+3 (bs-6951R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Sample:
heart (Mouse) Lysate at 40 ug
Primary: Anti-AKT1+2+3 (bs-6951R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 52 kD
Sample:
A431Cell (Human) Lysate at 30 ug
Hela Cell(Human)Lysate at 30 ug
Primary: Anti-AKT1+2+3(bs-6951R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (AKT1+2+3) Polyclonal Antibody, Unconjugated (bs-6951R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Tissue/cell: mouse embryo tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-AKT1+2+3 Polyclonal Antibody, Unconjugated(bs-6951R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-AKT1+2+3 Polyclonal Antibody, Unconjugated(bs-6951R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: Mouse embryos tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-AKT1 + 2 + 3 Polyclonal Antibody, Unconjugated(bs-6951R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control(blue): TM4 cells(fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody:Rabbit Anti-AKT1+2+3 antibody(bs-6951R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
 
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