Recombinant Human Active Akt1 Protein, CF

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Recombinant Human Active Akt1 Protein SDS-PAGE
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Product Details
Citations (4)
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Recombinant Human Active Akt1 Protein, CF Summary

Product Specifications

>70%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
The specific activity of Akt1 was determined to be 189 nmol/min/mg as per activity assay protocol.
Spodoptera frugiperda, Sf 9 (baculovirus)-derived human Akt1 protein
Accession #
N-terminal Sequence
Using an N terminal GST tag
85 kDa

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 10 mM glutathione, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: This product is stable at ≤ ‑70°C for up to 1 year from the date of receipt. For optimal storage, aliquot into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated freeze-thaw cycles.

Assay Procedure

  • Active Kinase - Active Akt1 (0.1 μg/μL) diluted with Kinase Dilution Buffer and assayed as outlined in sample activity plot. Note: These are suggested working dilutions. Optimal dilutions should be determined by each laboratory for each application.
  • Kinase Assay Buffer III (5X) - 200 mM Tris-HCl, pH 7.4, 100 mM MgCl2, and 0.5 mg/mL BSA. Add fresh DTT prior to use to a concentration of 50 μM.
  • Kinase Dilution Buffer IX (1X) - Kinase Assay Buffer III diluted at a 1:4 ratio (5x dilution) with cold water. Add fresh DTT to the aliquot prior to use to a final concentration of 50 μM.
  • ADP-Glo™ Kinase Assay Kit - ATP solution 10 mM, ADP solution 10 mM, ADP-Glo Reagent, Kinase Detection Reagent
  • Substrate - Akt (PKB) synthetic peptide substrate (CKRPRAASFAE) diluted in 25 mM Tris-HCl Buffer (pH 7.5) to a final concentration of 1 mg/mL.
  1. Thaw the Active Akt1, Kinase Assay Buffer III (5X), and Substrate on ice. Prepare a 15 μL enzyme dilution at the desired concentration, with Kinase Dilution Buffer IX (1X) in a pre-chilled 96-well plate.
  2. Prepare a Substrate/ATP mixture as follows (25μM example):
    a. 10 μM ATP solution: 1 μL
    b. Kinase Assay Buffer III (5X): 79 μL
    c. Substrate at 1 mg/mL: 80 μL
  3. Transfer the following reaction components prepared in step 2 to a 384-well opaque plate bringing the reaction volume up to 5 μL:
    a. 3 μL of diluted Active Akt1
    b. 2 μL of Substrate/ATP mix as prepared in step 2. This initiates the reaction.
  4. Set up the blank control as outlined in step 2, excluding the addition of the kinase. Replace the kinase with an equal volume of Kinase Dilution Buffer IX (1x).
  5. Incubate at ambient temperature for 40 minutes.
  6. After the 40 minute incubation period, terminate the reaction and deplete the remaining ATP by adding 5 μL of ADP-Glo™ Reagent. Spin down and shake the 384-well plate. Incubate the reaction mixture for another 40 minutes at ambient temperature.
  7. Add 10 μL of the Kinase Detection Reagent to the 384-well plate and incubate the reaction mixture for another 40 minutes at ambient temperature.
  8. Read the 384-well plate using the Luminescence Module Protocol on a GloMax®-Multi Microplate Multimode Reader.
  9. Determine the corrected activity (RLU) by removing the blank control value (see step 4) for each sample and calculate the kinase specific activity as outlined below.

    Calculation of Specific Activity of ADP (RLU/pmol)
    From ADP standard curve, determine RLU/pmol of ADP

    Kinase Specific Activity (SA) (pmol/minutes/μg or nmol/minutes/mg)
    Corrected RLU from reaction / [(SA of ADP in RLU/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)]

Scientific Data

SDS-PAGE Recombinant Human Active Akt1 Protein SDS-PAGE View Larger

The approximate molecular weight is 85 kDa and the purity is > 70%.

Reconstitution Calculator

Reconstitution Calculator

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Background: Akt1

Akt1, also known as PKB alpha, is a serine/threonine kinase that belongs to the Akt family. Akt1 is activated in cells in response to diverse stimuli such as hormones, growth factors and extracellular matrix components and is involved in glucose metabolism, transcription, survival, cell proliferation, angiogenesis, and cell motility (1). Akt1 is frequently over-expressed and active in many types of human cancers including cancers of colon, breast, brain, pancreas, and prostate as well as lymphomas and leukemias (2).

  1. Coffer, P.J. et al. (1998) Biochem J. 335:1.2.
  2. Anderson, K.E. et al. (1998) Curr. Biol. 8:684.
Long Name
v-Akt Murine Thymoma Viral Oncogene Homolog 1
Entrez Gene IDs
207 (Human); 11651 (Mouse); 25233 (Rat)
Alternate Names
AKT serine/threonine kinase 1; AKT; Akt1; AKT1m; CWS6; EC 2.7.11; EC; PKB alpha; PKB; PKBMGC99656; PRKBA; Protein Kinase B Alpha; Protein kinase B; Proto-oncogene c-Akt; rac protein kinase alpha; RAC; RAC-alpha serine/threonine-protein kinase; RAC-alpha; RAC-PK-alpha; RACPKB-ALPHA; Serine-Threonine Protein Kinase ; v-akt murine thymoma viral oncogene homolog 1; V-Akt Murine Thymoma Viral Oncogene-Like Protein 1

Citations for Recombinant Human Active Akt1 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. MPST deficiency promotes intestinal epithelial cell apoptosis and aggravates inflammatory bowel disease via AKT
    Authors: J Zhang, L Cen, X Zhang, C Tang, Y Chen, Y Zhang, M Yu, C Lu, M Li, S Li, B Lin, T Zhang, X Song, C Yu, H Wu, Z Shen
    Redox Biology, 2022-09-11;56(0):102469.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  2. UBE2S, a novel substrate of Akt1, associates with Ku70 and regulates DNA repair and glioblastoma multiforme resistance to chemotherapy
    Oncogene, 2016-09-05;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  3. The PI3K/Akt signal hyperactivates Eya1 via the SUMOylation pathway.
    Authors: Sun Y, Kaneko S, Li X, Li X
    Oncogene, 2014-06-23;34(19):2527-37.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Enzyme Assay
  4. Dithiolethione compounds inhibit Akt signaling in human breast and lung cancer cells by increasing PP2A activity.
    Authors: Switzer CH, Ridnour LA, Cheng RY, Sparatore A, Del Soldato P, Moody TW, Vitek MP, Roberts DD, Wink DA
    Oncogene, 2009-08-24;28(43):3837-46.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay


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Recombinant Human Active Akt1 Protein, CF
By Balaji Mahender on 01/09/2018
Application: Immunoassay Standard

Recombinant Human Active Akt1 Protein, CF
By Anonymous on 11/18/2017
Application: Immunoassay Standard