bs-5182R [Primary Antibody]
AKT1/2/3 (Thr305/308/309) Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: AKT1/2/3 (Thr305/308/309)

Specificity: The antibody recognizes phosphorylation at Thr308 of AKT1 and because of sequence similarity, it will also detect phosphorylation of Thr309 of AKT2 and phosphorylation at Thr305 of AKT3.

Modification Site: Thr305/308/309

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 207

Swiss Prot: Q9Y243

Source: KLH conjugated synthetic phosphopeptide derived from human AKT1 around the phosphorylation site of Thr308

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Shipped at 4°C. 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: 100ul

Concentration: 1ug/ul

Applications: WB(1:300-5000)
ELISA(1:500-1000)
FCM(1:20-100)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)

Predicted Molecular Weight: 53


Cross Reactive Species: Human
Mouse
Rat

Predicted Cross Reactive Species: Dog
Cow
Sheep
Pig
Chicken
Rabbit

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

PRODUCT SPECIFIC PUBLICATIONS
  • Wang Y et al. Aspirin inhibits adipogenesis of tendon stem cells and lipids accumulation in rat injury tendon through regulating PTEN/PI3K/AKT signalling. J Cell Mol Med. 2019 Nov;23(11):7535-7544.Read more>>
  • Zhou C et al. Combining transcatheter arterial embolization with iodized oil containing Apatinib inhibits HCC growth and metastasis. Sci Rep. 2020 Feb 19;10(1):2964. Read more>>
  • Zhou C et al. Combining transcatheter arterial embolization with iodized oil containing Apatinib inhibits HCC growth and metastasis. Sci Rep. 2020 Feb 19;10(1):2964. Read more>>
  • Li Hao. et al. Suppressive Role of Bam32/DAPP1 in Chemokine-Induced Neutrophil Recruitment. Int J Mol Sci. 2021 Jan;22(4):1825Read more>>
  • Dawoon Jeong. et al. ELOVL2: a novel tumor suppressor attenuating tamoxifen resistance in breast cancer. Am J Cancer Res. 2021; 11(6): 2568C2589Read more>>
  • ayarmaa Khadankhuu. et al. 10-O-(N N-Dimethylaminoethyl)-Ginkgolide B Methane-Sulfonate (XQ-1H) Ameliorates Cerebral Ischemia Via Suppressing Neuronal Apoptosis. J Stroke Cerebrovasc. 2021 Sep;30:105987Read more>>
  • Wen, Yukang. et al. Incomplete autophagy promotes the proliferation of Mycoplasma hyopneumoniae through the JNK and Akt pathways in porcine alveolar macrophages. VET RES. 2022 Dec;53(1):1-15Read more>>
  • Xiaoxiao Zhang. et al. Tanshinone A participates in the treatment of endometriosis by regulating adhesion, invasion, angiogenesis and inhibition of PI3K/Akt/mTOR signaling pathway. MOL MED REP. 2023 Nov;28(5):1-13Read more>>
VALIDATION IMAGES

Mouse lung lysates probed with Rabbit Anti-Akt1/2/3 (Thr305/308/309) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:300 overnight at 4˚C. Followed by a conjugated secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.


Formalin-fixed and paraffin embedded human breast cancer labeled with Rabbit Anti-AKT1/2/3 (Thr305/308/309) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:200 followed by conjugation to the secondary antibody and DAB staining


Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti-AKT1/2/3 (Thr305/308/309) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:200 followed by conjugation to the secondary antibody and DAB staining


Lane 1: Hela cell lysates; Lane 2: HepG2 cell lysates probed with phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C. The upper band could be the fully glycosylated form of the target protein.


Paraformaldehyde-fixed, paraffin embedded Mouse 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 (Thr305/308/309) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:400 overnight at 4°C, DAB staining.


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 (Thr305/308/309) Polyclonal Antibody, Unconjugated (bs-5182R) at 1:400 overnight at 4°C, DAB staining.


Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305)) polyclonal Antibody, Unconjugated (bs-5182R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.


Lane 1: Mouse NIH/3T3 cell lysates; Lane 2: Mouse Adrenal gland lysates; Lane 3: Human MCF-7 cell lysates; Lane 4: Human A549 cell lysates probed with AKT1/2/3 Polyclonal Antibody, Unconjugated (bs-5182R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.