Antigen Information

Gene Symbols:

Assay Format

Pathway:

MAPK Signaling
mTOR Signaling
NFkB Signaling
p53 Signaling
PKC 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
ATM (P-Ser1981)	eIF-2a (P-Ser52)	HDAC2 (P-Ser394)	HDAC4 (P-Ser632)	IKB-alpha (P-Ser32)
MSK1 (P-Ser376)	NF-kB (P-Ser536)	STAT1 (P-Ser727)	TAK1 (P-Ser412)	TBK1 (P-Ser172)
ZAP70 (P-Tyr292)

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 NF-kB Pathway 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 plastic Wrap
Distilled or de-ionized water
A chemiluminescent blot documentation system: CCD camera, X-ray Film and a suitable film processor, gel documentation system, or another chemiluminescent detection system capable of imaging a western blot.

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 - HeLa cells were grown to 80% confluency and then serum starved overnight. Cells were either untreated (bottom panel) or treated (top panel) with TNF alpha and Calyculin A. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

Note the strong signals of the Positive Control spots in the upper left and lower right corners. (See below for further details on the control spots.)

The signal intensity for each antigen-specific antibody spot is proportional to the relative concentration of the antigen in that sample. Comparison of signal intensities for individual antigen-specific antibody spots between and among array images can be used to determine relative differences in expression levels of each analyte sample-to-sample or group-to-group.

Human NF-κB Pathway Phosphorylation Array C1(图1)

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.