Proteome Profiler Human NFkB Pathway Array
Proteome Profiler Human NFkB Pathway Array 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 GuideKey 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.
- 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 |
Specifications
Product Datasheets
Data Example
Detection of Human NFkB Pathway Proteins in HCT-116 Cell Line. 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.
Detection of Human NFkB Pathway Proteins in Daudi Cell Line. 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).
Detection of Human NFkB Pathway Proteins in HT-29 Cell Line. 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).
Citations for Proteome Profiler Human NFkB Pathway Array
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.
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Citations: Showing 1 - 4
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Fasciola hepatica GST downregulates NF-?B pathway effectors and inflammatory cytokines while promoting survival in a mouse septic shock model
Authors: V Aguayo, BN Valdés Fer, M Rodríguez-, C Ruiz-Jimén, MJ Ramos-Bení, LB Méndez, AM Espino
Sci Rep, 2019;9(1):2275. 2019
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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
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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
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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
FAQs
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Do I need to use the protease inhibitors (e.g. aprotinin, Ipegal) if I only plan to use cell supernatants?
The proteinase inhibitors are recommended for cell and tissue lysates only. You do not need them for cell culture supernates.
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