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Human Insulin Receptor Phosphorylation Array C1 map

HepG2 or HeLa cells were grown to 80% confluency and then serum starved overnight. HepG2 Cells were treated with IGF for 10 minutes. HeLa Cells were treated with Calyculin A for 30 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

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Antigen Information

Gene Symbols:

EIF4E
FOXO3
IGF1R
INPP5D
INS
IRS1
PTPN11
SHC1
STK11

Assay Format

Pathway:

Akt Signaling
IGF Signaling
mTOR Signaling
PI3K-AKT Signaling

Number of Targets Detected:

Species Detected:

Human

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 (pg/ml)
Sandwich ELISA specificity
Higher density than ELISA, Western blot or bead-based multiplex

Target Names

Scroll over each target protein for more information
eIF-4E (P-Ser209) Gene Symbol: EIF4E EIF4EL1 EIF4F Gene ID: 1977 Accession #: P06730 ELISA Kit: RayBio Human Phospho-eIF-4E (S209) ELISA	FOXO-3 (P-Ser413) Gene Symbol: FOXO3 Gene ID: 2309 Accession #: O43524 ELISA Kit: RayBio Human and Mouse Phospho-FOXO3 (S253) ELISA	IGF-1 R (P-Tyr1165) Gene Symbol: IGF1R Gene ID: 3480 Accession #: P08069 ELISA Kit: RayBio Human Phospho-IGF1R (Y1165/1166)	Insulin (P-Tyr1189) Gene Symbol: INS Gene ID: 3630 Accession #: P01308 ELISA Kit: RayBio Human Phosphotyrosine Insulin R ELISA	IRS-1 (P-Ser318) Gene Symbol: IRS1 Gene ID: 3667 Accession #: P35568
LKB1 (P-Ser428) Gene Symbol: STK11 LKB1 PJS Gene ID: 6794 Accession #: Q15831 ELISA Kit: RayBio Human Phospho-LKB1 (Ser428) ELISA	SHC (P-Tyr427) Gene Symbol: SHC1 SHC SHCA Gene ID: 6464 Accession #: P29353 ELISA Kit: RayBio Human, Mouse and Rat Phospho-SHC (Y427) ELISA	SHIP1 (P-Tyr1020) Gene Symbol: INPP5D SHIP SHIP1 Gene ID: 3635 Accession #: Q92835 ELISA Kit: RayBio Human Phospho-SHIP1 (Y1020) ELISA	SHP-2 (P-Ser542) Gene Symbol: PTPN11 PTP2C SHPTP2 Gene ID: 5781 Accession #: Q06124 ELISA Kit: RayBio Custom Human SHP-2 ELISA

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 Insulin Receptor Phosphorylation Array C1 Membranes
Blocking Buffer
Detection Antibody Cocktail
1,000X HRP-Anti-Rabbit IgG Concentrate
20X Wash Buffer I Concentrate
20X Wash Buffer II Concentrate
2X Cell Lysis Buffer Concentrate
Detection Buffer C
Detection Buffer D
8-Well Incubation Tray w/ Lid
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 UVPs 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 - HepG2 or HeLa cells were grown to 80% confluency and then serum starved overnight. HepG2 Cells were treated with IGF for 10 minutes. HeLa Cells were treated with Calyculin A 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.

Manual

SDS

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