phospho-Akt (S473) Pan Specific Antibody (545007)

R&D Systems | Catalog # MAB887

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Simple Western

Cited:

Western Blot, Flow Cytometry, Infared Microscopy

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 545007
View all Akt Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human Akt (S473) site

Specificity

Detects human and mouse Akt1, Akt2 and Akt3, when phosphorylated at S473, S474 and S472, respectively.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for phospho-Akt (S473) Pan Specific Antibody (545007)

Detection of Human and Mouse Phospho-Akt (S473) antibody by Western Blot.

Detection of Human and Mouse Phospho-Akt (S473) by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF-7 human breast cancer cell line, and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 nm Calyculin for 30 minutes, 1 µg/mL insulin for 5 minutes, or 100 ng/mL Recombinant Human PDGF-AB (Catalog # 222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-Akt (S473) antibody by Simple WesternTM.

Detection of Human Phospho-Akt (S473) by Simple WesternTM.

Simple Western lane view shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human AKT by Simple Western

Detection of Human AKT by Simple Western

ACPA inhibits p-Akt, induces p-JNK and affects levels of specific metabolites in NSCLC lines.a Principal component analysis (PCA) score plot: Metabolomics profiling of control and ACPA-treated A549, H1299, H358, and H838 cells. b Changes in variable importance in projection (VIP) values for 19 metabolites in A549 cells. c, d, e Changes in VIP values for 20 metabolites in H1299, H358, and H838 cells. Significantly changed metabolites (*p < 0.05, indicated by arrows) were matched to apoptotic pathways. f, g, h, i Increase and decrease in several metabolites of ACPA-treated A549, H1299, H358, and H838 cells (*p < 0.05). j Simple Western showing total Akt, p-Akt (S473), total JNK46 and JNK54 and p-JNK46 and p-JNK54 (T183/Y185) in A549 cells at 24 hours after treatment with IC50 dose of ACPA. k Relative expression levels of Akt and p-Akt for control and ACPA-treated A549 cells after normalization by total vinculin protein. l Relative expression levels of JNK (46 and 54 kDa) and p-JNK for control and ACPA-treated A549 cells after normalization by total vinculin protein. *p < 0.05, Student’s t-test. All tests were done in quadruplicates. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33431819), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-Akt (S473) Pan Specific by Western Blot

Detection of Phospho-Akt (S473) Pan Specific by Western Blot

PTEN status in tested models (A) PTEN and pAKT levels in kidney cancer cell lines, IOD calculated relative to Vinculin (B) Relative to GusB expression of Pten in RNA isolates from kidney cancer and healthy kidney tissues (C) graphical workflow. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35625614), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-Akt (S473) Pan Specific by Western Blot

Detection of Phospho-Akt (S473) Pan Specific by Western Blot

PTEN status in tested models (A) PTEN and pAKT levels in kidney cancer cell lines, IOD calculated relative to Vinculin (B) Relative to GusB expression of Pten in RNA isolates from kidney cancer and healthy kidney tissues (C) graphical workflow. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35625614), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for phospho-Akt (S473) Pan Specific Antibody (545007)

Application
Recommended Usage

Simple Western

2 µg/mL
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1)

Western Blot

0.2 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, MCF‑7 human breast cancer cell line, and NIH‑3T3 mouse embryonic fibroblast cell line treated with calyculin, insulin, or Recombinant Human PDGF-AB (Catalog # 222-AB)

Reviewed Applications

Read 2 reviews rated 5 using MAB887 in the following applications:

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 PBS 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.

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Background: Akt

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.

Long Name

v-Akt Murine Thymoma Viral Oncogene Homolog

Alternate Names

PKB, RAC

Additional Akt Products

Product Documents for phospho-Akt (S473) Pan Specific Antibody (545007)

Certificate of Analysis

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Product Specific Notices for phospho-Akt (S473) Pan Specific Antibody (545007)

For research use only

Citations for phospho-Akt (S473) Pan Specific Antibody (545007)

Customer Reviews for phospho-Akt (S473) Pan Specific Antibody (545007) (2)

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  • Human/Mouse Phospho-Akt (S473) Pan Specific Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293 human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 07/21/2018
    phospho-Akt (S473) Pan Specific Antibody (545007) MAB887
  • Human/Mouse Phospho-Akt (S473) Pan Specific Antibody
    Name: Anonymous
    Application: Functional Assay
    Sample Tested: Purified Standard
    Species: Human
    Verified Customer | Posted 12/01/2017

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