Human Phospho-Akt1 (T308) Antibody

Name: Human Phospho-Akt1 (T308) Antibody
Brand: R&D Systems
SKU: MAB7419
Price: 459 USD
Availability: InStock

R&D Systems | Catalog # MAB7419

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Citations (11)

Product Documents

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Hamster - Mesocricetus auratus (Golden Hamster)

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₃ Clone # 658320

View all Akt1 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human Akt1 T308 site

Specificity

Detects human and mouse Akt1 when phosphorylated at T308.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₃

Scientific Data Images for Human Phospho-Akt1 (T308) Antibody

Detection of Human and Mouse Phospho-Akt1 (T308) by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-Akt1 (T308) in CCD‑1070Sk Human Cell Line.

Akt1 phosphorylated at T308 was detected in immersion fixed CCD-1070Sk human foreskin fibroblast cell line stimulated with Recombinant Human PDGF-BB (Catalog # 220-BB) using Mouse Anti-Human Phospho-Akt1 (T308) Mono-clonal Antibody (Catalog # MAB7419) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human AKT1 by Western Blot

ASPP2 blocks Gal-1 dependent nanoclustering and halts oncogenic H-ras induced transformation.Nanoclustering-FRET analysis in HEK cells coexpressing mGFP- and mCherry-tagged (A, B) H-rasG12V or (C, D) K-rasG12V. Cells were analysed after overexpression of either Gal-1 or ASPP2 plasmids, or both (1:1 ratio). Plotted are the means ± SEM, n = 3. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left) or K-rasG12V (right) alone or together with Gal-1 and ASPP2. Statistical significance of differences was examined using t-test (n = 3; *, p<0.05, **, p< 0.01, ***, p<0.001). (F, G) Colony survival assay of NIH/3T3 cells stably expressing (F) H-rasG12V or (G) K-rasG12V and transiently expressing indicated constructs. Colony survival was graphed based on mean foci areas calculated from at least 4 independent biological repeats. (A-D, F-G) Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (ns, not significant; *, p<0.05; **, p<0.01; ****, p<0.0001). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human AKT1 by Western Blot

ASPP2 increases oncogenic H-ras, K-ras and N-ras-effector-recruitment, as well as ERK- and AKT-signalling.(A) Left, scheme explaining effector-recruitment FRET analysis in HEK cells. Right, examples of FLIM-FRET images of HEK cells from the different FRET samples as indicated. (B-D) Effector-recruitment FRET analysis in HEK cells coexpressing (B) mGFP-H-rasG12V, (C) mGFP-K-rasG12V or (D) mGFP-NrasG12V and mRFP-RBD from C-Raf. The effect of Gal-1 or ASPP2 expression on effector-recruitment FRET was compared to control samples. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left), K-rasG12V (middle) or N-rasG12V (right) without or with Gal-1 or ASPP2. Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (mean ± SEM, n = 3; ns, not significant; *, p<0.05, **, p< 0.01, ***, p<0.001, ****, p< 0.0001). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human Phospho-Akt1 (T308) Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed CCD‑1070Sk human foreskin fibroblast cell line stimulated with Recombinant Human PDGF‑BB (Catalog # 220-BB)

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and NIH‑3T3 mouse embryonic fibroblast cell line treated with Recombinant Human PDGF‑BB (Catalog # 220-BB) or Calyculin A

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in Tris with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: Akt1

Akt, also known as protein kinase B (PKB), is a central kinase in such diverse cellular processes as glucose uptake, cell cycle progression, and apoptosis. Three highly homologous members define the Akt family: Akt1 (PKB alpha ), Akt2 (PKB beta ), and Akt3 (PKB gamma ). All three Akts contain an amino-terminal pleckstrin homology domain, a central kinase domain, and a carboxyl-terminal regulatory domain. Akt1 is the most widely expressed family member and is frequently activated in a number of carcinomas, including breast, prostate, lung, pancreatic, liver, ovarian, and colorectal cancer. Akt1 is activated in a multistep process that involves the sequential phosphorylation of Thr450 by JNK kinases, Thr308 by PDK1, and Ser473 by PDK2 or mTORC2. Activated Akt1 phosphorylates a wide variety of cytosolic, nuclear, and mitochondrial substrates. Human Akt1 shares 98% aa sequence identity with mouse and rat Akt1. MAB7419 also detects mouse Phospho-Akt1 (T308) in Western Blot.