Gene Symbols:ATF2, FOS, JUN, MAP3K7, SMAD1, SMAD2, SMAD4, SMAD5
Pathway:PKC Signaling
TGF-beta Signaling
Number of Targets Detected:8
Method of Detection:Chemiluminescence
Quantitative/Semi-Quantitative:Semi-Quantitative
Size:2, 4, or 8 Sample Kit
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
| Scroll over each target protein for more information |
|---|
c-Jun (P-Ser73) | SMAD1 (P-Ser463/465) | SMAD5 (P-Ser463/465) | SMAD4 (P-Thr277) | ATF2 (P-Thr69/71) |
C-Fos (P-Thr232) | SMAD2 (P-Ser245/250/255) | TAK1 (P-Ser412) |
|
|
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 TGF beta Phosphorylation Array C1 Membranes
Blocking Buffer
Detection Antibody Cocktail
500X 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 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
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 Calyculin A for 30 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.
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.