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.

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

Each antibody is supplied as a 50X stock; enough for 25 assays when used in 500 µL staining volume per assay. Each 1X antibody solution has an endotoxin level of < 5 EU/mL.

Stability and Storage

For a complete list of the kit contents and necessary materials, please see the Materials Provided/Other Supplies Required sections of the product datasheet.

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) Cell-based ELISA Kits
ASC          
BCL-10          
CARD6          
CD40/TNFRSF5 DCCD40 DY632-05      
dAP1/BIRC2          
dAP2/BIRC3          
FADD/MORT1          
Fas/TNFRSF6/CD95 DFS00 DY326      
IκBα     DYC4299-2   KCB4809
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         KCB7226
RelΑ/p65 (pS529)          
c-Rel          
SHARPIN          
SOCS-6          
STAT1p91          
STAT1 (pY701)         KCB2894
STAT2          
STAT2 (pY689)         KCB2890
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.
View Larger Image

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).

 

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

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