Recombinant Human Active Akt1 Protein, CF

R&D Systems | Catalog # 1775-KS

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems Sf 9 (baculovirus)-derived Recombinant Human Active Akt1 Protein (1775-KS)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 9 (baculovirus)

Accession Number

NM_005163

Applications

Bioactivity
View all Akt1 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 9 (baculovirus)-derived human Akt1 protein

Purity

>70%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

N-terminal Sequence Analysis

Using an N terminal GST tag

SDS-PAGE

85 kDa

Activity

The specific activity of Akt1 was determined to be 189 nmol/min/mg as per activity assay protocol.

Reviewed Applications

Read 2 reviews rated 4 using 1775-KS in the following applications:

Scientific Data Images for Recombinant Human Active Akt1 Protein, CF

Recombinant Human Active Akt1 Protein SDS-PAGE

Recombinant Human Active Akt1 Protein SDS-PAGE

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

Formulation, Preparation, and Storage

1775-KS
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.

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).

References

  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

Alternate Names

PKB alpha, PRKBA, RAC-alpha

Entrez Gene IDs

207 (Human); 11651 (Mouse); 25233 (Rat)

Gene Symbol

AKT1

UniProt

NM_005163 (Human)

Additional Akt1 Products

Product Documents for Recombinant Human Active Akt1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Active Akt1 Protein, CF

For research use only

Citations for Recombinant Human Active Akt1 Protein, CF

Customer Reviews for Recombinant Human Active Akt1 Protein, CF (2)

4 out of 5
2 Customer Ratings
5 Stars
50%
4 Stars
0%
3 Stars
50%
2 Stars
0%
1 Stars
0%

Have you used Recombinant Human Active Akt1 Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card
Showing  1 - 2 of 2 reviews Showing All
Filter By:
Clear Filters X
  • Name: Balaji Mahender
    Application: Immunoassay Standard
    Verified Customer | Posted 01/09/2018
  • Recombinant Human Active Akt1 Protein, CF
    Name: Anonymous
    Application: Immunoassay Standard
    Verified Customer | Posted 11/18/2017

There are no reviews that match your criteria.

Showing  1 - 2 of 2 reviews Showing All

Protocols

View specific protocols for Recombinant Human Active Akt1 Protein, CF (1775-KS):

Materials
  • 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)]

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes