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Western Blot (WB) (Figure 1. Western blot analysis of AKT2 using anti-AKT2 antibody (M00725-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human A549 whole cell lysate, Lane 2: human 293T whole cell lysate, Lane 3: human HELA whole cell lysate, Lane 4: human Caco-2 whole cell lysate, Lane 5: human K562 whole cell lysate, Lane 6: human HL-60 whole cell lysate, Lane 7: human PC-3 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AKT2 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )

Mouse anti-Human AKT2 Monoclonal Antibody | anti-AKT2 antibody

Anti-AKT2 Antibody (monoclonal, 10C6)

Gene Names
AKT2; PKBB; PRKBB; HIHGHH; PKBBETA; RAC-BETA
Reactivity
Human
Applications
Western Blot, Immunocytochemistry
Purity
Immunogen Affinity Purified
Synonyms
AKT2; Monoclonal Antibody; Anti-AKT2 Antibody (monoclonal; 10C6); RAC-beta serine/threonine-protein kinase; Protein kinase Akt-2; Protein kinase B beta; PKB beta; RAC-PK-beta; V-akt murine thymoma viral oncogene homolog 2; anti-AKT2 antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
IgG1
Clone Number
10C6
Specificity
No cross reactivity with other proteins.
Purity/Purification
Immunogen Affinity Purified
Form/Format
Lyophilized
Sequence Length
438
Applicable Applications for anti-AKT2 antibody
Western Blot (WB), Immunocytochemistry (ICC)
Application Notes
WB: 0.1-0.5 mug/ml
ICC: 0.5-1 mug/ml
Tested Species: In-house tested species with positive results.
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human AKT2 (454-481aa DRYDSLGLLELDQRTHFPQFSYSASIRE), different from the related mouse sequence by two amino acids, and from the related rat sequence by one amino acid.
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Relevant Detection Systems
It is recommended to use an Enhanced Chemiluminescent Kit with anti-Rabbit IgG (MBS176460) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (MBS176453) for ICC.
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time.
Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of AKT2 using anti-AKT2 antibody (M00725-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human A549 whole cell lysate, Lane 2: human 293T whole cell lysate, Lane 3: human HELA whole cell lysate, Lane 4: human Caco-2 whole cell lysate, Lane 5: human K562 whole cell lysate, Lane 6: human HL-60 whole cell lysate, Lane 7: human PC-3 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AKT2 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )

Western Blot (WB) (Figure 1. Western blot analysis of AKT2 using anti-AKT2 antibody (M00725-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human A549 whole cell lysate, Lane 2: human 293T whole cell lysate, Lane 3: human HELA whole cell lysate, Lane 4: human Caco-2 whole cell lysate, Lane 5: human K562 whole cell lysate, Lane 6: human HL-60 whole cell lysate, Lane 7: human PC-3 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AKT2 antigen affinity purified monoclonal antibody at 0.5 ug/ml overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. )
Related Product Information for anti-AKT2 antibody
Description: Mouse IgG monoclonal antibody for AKT2 detection. Tested with WB, ICC in Human.
Background: AKT2 is a putative oncogene encoding a protein belonging to a subfamily of serine/threonine kinases containing SH2-like (Src homology 2-like) domains. This gene is mapped to 19q13.2. AKT2 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. AKT2 seems also to be the principal isoform responsible of the regulation of glucose uptake. AKT2 is also specifically involved in skeletal muscle differentiation, one of its substrates in this process being ANKRD2. Overexpression of AKT2 contributes to the malignant phenotype of a subset of human ductal pancreatic cancers.
References
1. Cheng, J. Q., Ruggeri, B., Klein, W. M., Sonoda, G., Altomare, D. A., Watson, D. K., Testa, J. R. Amplification of AKT2 in human pancreatic cancer cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA. Proc. Nat. Acad. Sci. 93: 3636-3641, 1996. 2. Li, X., Monks, B., Ge, Q., Birnbaum, M. J. Akt/PKB regulates hepatic metabolism by directly inhibiting PGC-1-alpha transcription coactivator. Nature 447: 1012-1016, 2007.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
208
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
51,083 Da
NCBI Official Full Name
RAC-beta serine/threonine-protein kinase isoform 4
NCBI Official Synonym Full Names
AKT serine/threonine kinase 2
NCBI Official Symbol
AKT2
NCBI Official Synonym Symbols
PKBB; PRKBB; HIHGHH; PKBBETA; RAC-BETA
NCBI Protein Information
RAC-beta serine/threonine-protein kinase
UniProt Protein Name
RAC-beta serine/threonine-protein kinase
Protein Family
UniProt Gene Name
AKT2
UniProt Synonym Gene Names
PKB beta

NCBI Description

This gene is a putative oncogene encoding a protein belonging to a subfamily of serine/threonine kinases containing SH2-like (Src homology 2-like) domains. The gene was shown to be amplified and overexpressed in 2 of 8 ovarian carcinoma cell lines and 2 of 15 primary ovarian tumors. Overexpression contributes to the malignant phenotype of a subset of human ductal pancreatic cancers. The encoded protein is a general protein kinase capable of phophorylating several known proteins. [provided by RefSeq, Jul 2008]

Uniprot Description

AKT2 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI3P-5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI3K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development.One of the few specific substrates of AKT2 identified recently is PITX2. Phosphorylation of PITX2 impairs its association with the CCND1 mRNA-stabilizing complex thus shortening the half-life of CCND1. AKT2 seems also to be the principal isoform responsible of the regulation of glucose uptake. Phosphorylates C2CD5 on 'Ser-197' during insulin-stimulated adipocytes. AKT2 is also specifically involved in skeletal muscle differentiation, one of its substrates in this process being ANKRD2. Down-regulation by RNA interference reduces the expression of the phosphorylated form of BAD, resulting in the induction of caspase-dependent apoptosis. Phosphorylates CLK2 on 'Thr-343'.CautionIn light of strong homologies in the primary amino acid sequence, the 3 AKT kinases were long surmised to play redundant and overlapping roles. More recent studies has brought into question the redundancy within AKT kinase isoforms and instead pointed to isoform specific functions in different cellular events and diseases. AKT1 is more specifically involved in cellular survival pathways, by inhibiting apoptotic processes; whereas AKT2 is more specific for the insulin receptor signaling pathway. Moreover, while AKT1 and AKT2 are often implicated in many aspects of cellular transformation, the 2 isoforms act in a complementary opposing manner. The role of AKT3 is less clear, though it appears to be predominantly expressed in brain.

Research Articles on AKT2

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Product Notes

The AKT2 akt2 (Catalog #AAA1752014) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-AKT2 Antibody (monoclonal, 10C6) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's AKT2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC). WB: 0.1-0.5 mug/ml ICC: 0.5-1 mug/ml Tested Species: In-house tested species with positive results. Researchers should empirically determine the suitability of the AKT2 akt2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "AKT2, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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