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Human/Mouse AKT Pathway Phosphorylation Array C1

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RayBio® C-Series Human/Mouse AKT Pathway Phosphorylation Array C1. Detects 18 phosphorylated human and mouse proteins. Suitable for all liquid sample types but intended for use with cell and tissue lysates.

$752.97  
AAH-AKT-1-8
+ −

Assay Format

Pathway:
  • Akt Signaling
  • AMPK Signaling
  • HER/ErbB Signaling
  • HIF-1 alpha Signaling
  • IGF Signaling
  • JAK/STAT Signaling
  • MAPK Signaling
  • mTOR Signaling
  • PI3K-AKT Signaling
  • TGF-beta Signaling
  • Wnt/beta-Catenin Signaling
Number of Targets Detected:
18
Species Detected:
  • Human
  • Mouse
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
Chemiluminescence
Quantitative/Semi-Quantitative:
  • Semi-Quantitative
Solid Support:
Membrane

Product Specifications

Size:
2, 4, or 8 Sample Kit

Protein Information

Gene Symbols:
AKT1 | AKT1S1 | BAD | CDKN1B | EIF4EBP1 | GSK3A | GSK3B | MAPK1 | MAPK3 | MTOR   View more

Product Features

  • Easy to use
  • No specialized equipment needed
  • Compatible with nearly any liquid sample
  • Proven technology (many publications)
  • Highly sensitive
  • Sandwich ELISA specificity
  • Higher density than ELISA, Western blot or bead-based multiplex

Target Names

Scroll over each target protein for more information
Akt (P-Ser473)
(PKB (P-Ser473))
AMPKa (P-Thr172)
  • Gene Symbol: PRKAA1 AMPK1
  • Gene ID: 5562
  • Accession #: Q13131
BAD (P-Ser112)
4E-BP1 (P-Thr36)
ERK1 (P-T202/Y204)/ERK2 (P-Y185/Y187)
GSK3a (P-Ser21)
GSK3b (P-Ser9)
mTOR (P-Ser2448)
p27 (P-Thr198)
P53 (P-Ser15)
P70S6K (P-Thr421/Ser424)
PDK1 (P-Ser241)
  • Gene Symbol: PDK1 PDHK1
  • Gene ID: 5170
  • Accession #: O15530
PRAS40 (P-Thr246)
PTEN (P-Ser380)
Raf-1 (Ser301)
RPS6 (P-Ser235/Ser236)
RSK1 (P-Ser380)
RSK2 (P-Ser386)
  • Gene Symbol: RPS6KA3 ISPK1 MAPKAPK1B RSK2
  • Gene ID: 10432
  • Accession #: P51812

Application Notes

Suggested Application
Multiplexed Protein Detection; Detection of Relative Protein Expression; Detecting Patterns of Cytokine Expression; Biomarker/ Key Factor Screening; Identifying Key Factors; Confirming a Biological Process
Kit Components
  • Human and Mouse AKT Pathway Phosphorylation Antibody Array C1 Membranes
  • Blocking Buffer
  • Detection Antibody Cocktail
  • 1,000X HRP-Anti-Rabbit IgG Concentrate
  • 20X Wash Buffer I Concentrate
  • 20X Wash Buffer II Concentate
  • 2X Cell Lysis Buffer Concentrate
  • Detection Buffer C
  • Detection Buffer D
  • 8-Well Incubation Tray
  • Protease Inhibitor Cocktail
  • 100x Phosphatase Inhibitor Cocktail I
  • Phosphatase Inhibitor Cocktail II
  • Plastic Sheets
  • Array Map Template
  • Manual
Other Materials Required
  • Pipettors, pipet tips and other common lab consumables
  • Orbital shaker or oscillating rocker
  • Tissue Paper, blotting paper or chromatography paper
  • Adhesive tape or Saran Wrap
  • Distilled or de-ionized water
  • A chemiluminescent blot documentation system (such as UVP's ChemiDoc-It® or EpiChem II Benchtop Darkroom or GE's ImageQuant™ LAS 4000 or Amersham Imagers 600 and 680), X-ray Film and a suitable film processor, or another chemiluminescent detection system.
Protocol Outline
  1. Block membranes
  2. Incubate with Sample
  3. Incubate with Detection Antibody Cocktail
  4. Incubate with HRP-Conjugated anti-IgG
  5. Incubate with Detection Buffers
  6. Image with chemiluminescent imaging system
  7. Perform densitometry and analysis

Typical Data

Figure 1 - NIH3T3 cells were grown to 80% confluency and then serum starved overnight. Cells were either untreated (bottom panel) or treated (top panel) with PDGFβ for 10 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.
Figure 2 - 293 cells were grown to 80% confluency and then serum starved 24 hours. Cells were either untreated (bottom panel) or treated (top panel) with Insulin for 30 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

Storage/Stability

For best results, store the entire kit frozen at -20°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array membranes and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months.
  1. Qiao L, Hu S, Liu S, et al. microRNA-21-5p dysregulation in exosomes derived from heart failure patients impairs regenerative potential. J Clin Invest. 2019;129(6):2237-2250. Published 2019 Apr 29. doi:10.1172/JCI123135
    Species: Human
    Sample type: Cell Lysate (iCM, H9C2, HCF, HUVEC cells )
  2. Yang Y, Wang J, Guo S, et al. Non-lethal sonodynamic therapy facilitates the M1-to-M2 transition in advanced atherosclerotic plaques via activating the ROS-AMPK-mTORC1-autophagy pathway. Redox Biol. 2020;32:101501. doi:10.1016/j.redox.2020.101501
    Species: Mouse
    Sample type: Conditioned Media
  3. Luszczak, S., Simpson, B.S., Stopka-Farooqui, U. et al. Co-targeting PIM and PI3K/mTOR using multikinase inhibitor AUM302 and a combination of AZD-1208 and BEZ235 in prostate cancer. Sci Rep 10, 14380 (2020). https://doi.org/10.1038/s41598-020-71263-9
    Species: Human, Mouse
    Sample type: Conditioned media (cell culture media)
  4. Luszczak S, Simpson BS, Stopka-Farooqui U, Sathyadevan VK, Echeverria LMC, Kumar C, Costa H, Haider A, Freeman A, Jameson C, Ratynska M, Ben-Salha I, Sridhar A, Shaw G, Kelly JD, Pye H, Gately KA, Whitaker HC, Heavey S. Co-targeting PIM and PI3K/mTOR using multikinase inhibitor AUM302 and a combination of AZD-1208 and BEZ235 in prostate cancer. Sci Rep. 2020 Sep 1;10(1):14380. doi: 10.1038/s41598-020-71263-9. Erratum in: Sci Rep. 2020 Nov 11;10(1):19870. PMID: 32873828; PMCID: PMC7463239.
    Species: Human
    Sample type: Conditioned Media
  5. Chiurchiu V., Rapino C., Talamonti E., et al. Anandamide Suppresses Proinflammatory T Cell Responses In Vitro through Type-1 Cannabinoid Receptor-Mediated mTOR Inhibition in Human Keratinocytes. J Immunol. 2016 Nov 1;197(9):3545-3553. doi: 10.4049/jimmunol.1500546
    Species: Human
    Sample type: Cell Lysate (Human keratinocytes and kidney epidermal cells treated with anandamide)
  6. Chiurchiù, Valerio, et al. "Anandamide suppresses proinflammatory T cell responses in vitro through type-1 cannabinoid receptor–mediated mTOR inhibition in human keratinocytes." The Journal of Immunology 197.9 (2016): 3545-3553.
    Species: Human
    Sample type: Cell Lysate
  7. Lu J, Liu QH, Wang F, et al. Exosomal miR-9 inhibits angiogenesis by targeting MDK and regulating PDK/AKT pathway in nasopharyngeal carcinoma. J Exp Clin Cancer Res. 2018;37(1):147. Published 2018 Jul 13. doi:10.1186/s13046-018-0814-3
    Species: Human
    Sample type: Cell Lysate
  8. Li M, Yang J, Zhang L, et al. A low-molecular-weight compound exerts anticancer activity against breast and lung cancers by disrupting EGFR/Eps8 complex formation. J Exp Clin Cancer Res. 2019;38(1):211. Published 2019 May 22. doi:10.1186/s13046-019-1207-y
    Species: Human
    Sample type: Cell Lysate (MDA-MB-468 )
  9. Cai S, Yang Q, Cao Y, et al. PF4 antagonizes retinal neovascularization via inhibiting PRAS40 phosphorylation in a mouse model of oxygen-induced retinopathy. Biochim Biophys Acta Mol Basis Dis. 2020;1866(3):165604. doi:10.1016/j.bbadis.2019.165604
    Species: Mouse
    Sample type: Vitreous Fluid (retinal fluid)
Worked fine as expected with minimal background. Great product when you have smaller sample size.
Laxmi
WUSTL 07/22/2022, 05:28 PM
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