Proteome Profiler Human NFkB Pathway Array

Contains 4 membranes-each spotted in duplicate with 41 different proteins and 4 serine or tyrosine phosphorylation sites antibodies
  (3 citations)     
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Product Details
Citations (3)
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Kit Summary

A membrane-based antibody array for the parallel determination of the relative levels of selected human NFkB Pathway proteins. Validated for analyte detection in cell lysates.

Troubleshooting Guide

Key Benefits

  • Detects 41 human proteins and 4 serine or tyrosine phosphorylation sites simultaneously
  • Requires no specialized equipment
 

 

Principle of the Assay

Carefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell lysates are diluted and mixed with a cocktail of biotinylated detection antibodies. The sample/antibody mixture is then incubated with the array. Any protein/detection antibody complex present is bound by its cognate immobilized capture antibody on the membrane. Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of protein bound. Chemiluminescence is detected in the same manner as a Western blot.

Why Use an Antibody Array to Detect Multiple Proteins?

Determining the expression of multiple Proteins in a single sample can be expensive, time consuming and can require specialized equipment. Performing multiple immunoprecipitations and Western blots requires time, labor, and reagents. The use of a multiplex antibody array to detect multiple proteins in a single sample can be cost-effective and also save time and sample.

 

 

Kit Contents
  • 4 Array Membranes
  • 4-Well Multi-dish
  • Array Buffers
  • Wash Buffer
  • Detection Antibody Cocktail, Human NF kappa B Pathway Array
  • Streptavidin-HRP
  • Chemiluminescent Detection Reagents
  • Transparency Overlay Template
  • Detailed Protocol

Stability and Storage

Reagents are stable for 12 months from date of receipt when stored in the dark at 2° C to 8° C.
 

Simultaneously detect the levels of these proteins in a single sample.
ASC IL-18 R alpha RelA/p65 (pS529)
BCL-10 IRAK1 c-Rel
CARD6 IRF5 SHARPIN
CD40/TNFRSF5 IRF8 SOCS-6
dAP1/BIRC2 JNK1/2 STAT1p91
dAP2/BIRC3 JNK2 STAT1 (pY701)
FADD/MORT1 LTBR/TNFRSF3 STAT2
Fas/TNFRSF6/CD95 Metadherin/AEG-1 STAT2 (pY689)
IκBα MYD88 STING/TMEM173
IκBε NFκB1 TLR2
IKK1/IKKα/CHUK NFκB2 TNF RI/TNFRSF1A
IKK2/IKKβ NGF R/TNFRSF16 TNF RII/TNFRSF1B
IKKυ/NEMO P53 TRAF2
IL-1 RI P53 (pS46) TRAIL R1/DR4
IL-17 RA RelA/p65 TRAIL R2/DR5
 

 

Analyte Quantikine® ELISA Kits DuoSet® ELISA Development System DuoSet® IC ELISA Development System (Total) DuoSet® IC ELISA Development System (Phospho)
ASC        
BCL-10        
CARD6        
CD40/TNFRSF5 DCCD40 DY632-05    
dAP1/BIRC2        
dAP2/BIRC3        
FADD/MORT1        
Fas/TNFRSF6/CD95 DFS00 DY326    
IκBα     DYC4299-2  
I&kappaBε        
IKK1/IKKα/CHUK        
IKK2/IKKβ        
IKKΥ/NEMO        
IL-1 RI   DY269    
IL-17 RA   DY177    
IL-18 Rα        
IRAK1        
IRF5        
IRF8        
JNK1/2     DYC1205-2 DYC1387-2
JNK2     DYC1846-2 DYC2236-2
LTBR/TNFRSF3        
Metadherin/AEG-1        
MyD88   DY2928    
NFκB1        
NFκB2        
NGF R/TNFRSF16   DY367    
P53     DYC1043-2  
P53 (pS46)       DYC1489-2
RelΑ/p65        
RelΑ/p65 (pS529)        
c-Rel        
SHARPIN        
SOCS-6        
STAT1p91        
STAT1 (pY701)        
STAT2        
STAT2 (pY689)        
STING/TMEM173        
TLR2 DTLR20 DY2616    
TNF RI/TNFRSF1A DRT100 DY225    
TNF RII/TNFRSF1B DRT200 DY726    
TRAF2        
TRAIL R1/DR4        
TRAIL R2/DR5        

 

 

 

Data Examples
Human Induced Pluripotent Stem Cells Express TRA-1-81.
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HCT-116 Untreated vs. Epoxomicin Treated
HCT-116 human colorectal carcinoma cells were untreated or treated with 1 µM Epoxomicin (Boston Biochem, Catalog # I-110) for 6 hours (500 µg lysate, 5 minute exposure). RS = Reference Spots.

Daudi Untreated vs. IFN-alpha 2A Treated
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Daudi Untreated vs. IFN-alpha 2A Treated
Daudi human Burkitt’s lymphoma cells were untreated or treated with 500 U/mL IFN-alpha 2A (R&D Systems, Catalog # 11100-1) for five minutes (1 mg lysate, 10 minute exposure).

HT-29 Untreated vs. TNF-alpha +/- MLN4924 Treatment
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HT-29 Untreated vs. TNF-alpha +/- MLN4924 Treatment
HT-29 human colorectal carcinoma cells were untreated or treated with 50 ng/mL recombinant human TNF-alpha (R&D Systems, Catalog # 210-TA) for ten minutes with or without pretreatment with 5 µM MLN4924 (Boston Biochem, Catalog # I-502) for 1 hour (400 µg lysate, 5 minute exposure).

 


Product Datasheets

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Preparation and Storage
  • Stability & Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Sample Type
  1. Cardiac glycosides suppress the maintenance of stemness and malignancy via inhibiting HIF-1? in human glioma stem cells
    Authors: DH Lee, S Cheul Oh, AJ Giles, J Jung, MR Gilbert, DM Park
    Oncotarget, 2017;8(25):40233-40245.  2017
  2. Adrenomedullin Suppresses Vascular Endothelial Growth Factor-Induced Vascular Hyperpermeability and Inflammation in Retinopathy
    Authors: A Imai, Y Toriyama, Y Iesato, K Hirabayash, T Sakurai, A Kamiyoshi, Y Ichikawa-S, H Kawate, M Tanaka, T Liu, X Xian, L Zhai, K Dai, K Tanimura, T Liu, N Cui, A Yamauchi, T Murata, T Shindo
    Am. J. Pathol, 2017;0(0):.  2017
  3. Calpain inhibition decreases inflammatory protein expression in vessel walls in a model of chronic myocardial ischemia
    Authors: Brittany A Potz
    Surgery, 2016;0(0):.  2016

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