Human NF-κB Pathway Phosphorylation Array C1

Arrays|Post-Translational Modification Arrays|Phosphorylation Arrays|Antibody Arrays|Membrane Antibody Arrays

-20°C

Cell Culture Supernatants
Cell Lysates
Plasma
Serum
Tissue Lysates

Chemiluminescence

Membrane

Human NF-kB Pathway Phosphorylation Array C1 Membranes
Blocking Buffer
Detection Antibody Cocktail1, 000
X HRP-Anti-Rabbit-Ig
G Concentrate20
X Wash Buffer I Concentrate20
X Wash Buffer I
I Concentrate2
X Cell Lysis Buffer Concentrate
Detection Buffer C
Detection Buffer D8-Well Incubation Tray w/ Lid
Protease Inhibitor Cocktail100x Phosphatase Inhibitor Cocktail I
Phosphatase Inhibitor Cocktail IIPlastic Sheets
Array Map Template
Manual

Block membranes
Incubate with Sample
Incubate with Detection Antibody Cocktail
Incubate with HRP-Conjugated anti-IgGIncubate with Detection Buffers
Image with chemiluminescent imaging system
Perform densitometry and analysis

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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.