Antigen Information

Gene Symbols:

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:

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

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)	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)	PRAS40 (P-Thr246)	PTEN (P-Ser380)	Raf-1 (Ser301)
RPS6 (P-Ser235/Ser236)	RSK1 (P-Ser380)	RSK2 (P-Ser386)

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

Block membranes
Incubate with Sample
Incubate with Detection Antibody Cocktail
Incubate with HRP-Conjugated anti-IgG
Incubate with Detection Buffers
Image with chemiluminescent imaging system
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.
Human/Mouse AKT Pathway Phosphorylation Array C1(图1)

Human/Mouse AKT Pathway Phosphorylation Array C1(图1)

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.
Human/Mouse AKT Pathway Phosphorylation Array C1(图2)

Human/Mouse AKT Pathway Phosphorylation Array C1(图2)

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.