NF kappaB p65 (pS536) ELISA Kit is a single-wash 90-min Simplestep used to quantify NF kappaB p65 (pS536) with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 15 citations
View Alternative Names
NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3
- sELISA
Supplier Data
Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of NFκB p65 (pS536) are normalized and plotted.
- sELISA
Supplier Data
Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
Typical cell lysate dilution series.
Example of a typical NFκB p65 (pS536) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
- sELISA
Supplier Data
Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
Comparison of NF kappaB p65 (pS536) expression in different cell lines.
Cell line analysis for Total NFκB p65 from 200 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
- sELISA
Supplier Data
Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
NF kappaB p65 (pS536) phosphorylation in response to TNFα treatment.
Induction of NFκB p65 (pS536) phosphorylation in MCF-7 cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of NFκB p65 (pS536) are plotted and compared against total NFκB p65 protein levels. Comparative NFκB p65 (pS536) and NFκB p65 (Total) data also shown by Western Blot.
Reactivity data
Product details
Abcam's NFκB p65 (pS536) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFκB p65 (pS536) protein in Human and mouse cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Precision
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Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.
Pathways
NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.
Product protocols
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Publications (22)
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Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics 18:2664-2681 PubMed34664178
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Molecular medicine reports 23: PubMed33537800
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Scientific reports 10:14693 PubMed32895407
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Product promise
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