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 - 5
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TNFa and IL2 Encoding Oncolytic Adenovirus Activates Pathogen and Danger-Associated Immunological Signaling
Authors: C Heiniö, R Havunen, J Santos, K Lint, V Cervera-Ca, A Kanerva, A Hemminki
Cells, 2020;9(4):. 2020
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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.
Reviews for Proteome Profiler Human NFkB Pathway Array
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