Abcam, ab176663, NF kappaB p65 (pS536 + Total) ELISA Kit

Size: 1 x 96Tests
NF kappaB p65 (pS536 + Total) ELISA Kit is a single-wash 90-min Simplestep used to detect NF kappaB p65 (pS536 + Total). The assay uses a simple mix-wash-read protocol with just one incubation and one wash step. - Colorimetric Semi-quantitative Sandwich ELISA - 450 nm readout : works on any standard plate reader - Cited in over 25 citations
Key facts
Detection method:Colorimetric,
Sample types:Tissue Homogenate, Cell Lysate,
Reacts with:Mouse, Human,
Assay type:Semi-quantitative,
Assay time:1h 30m,
Assay Platform:Microplate

Product details:
Abcam's NFkB p65 (pS536) and NFkB p65 (Total)
in vitro
SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFkB p65 (pS536) and Total NFkB p65 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.
Estimated sensitivity
: Phospho-NF-κB p65 (Ser536): 5 μg/mL (tested with HeLa), Total NF-κB p65: 5 μg/mL (tested with HeLa)
: Phospho-NF-κB p65 (Ser536): 6-600 μg/mL, Total NF-κB p65: 6-600 μg/mL
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.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
NF-kB p65 also known as RelA is a transcription factor with a molecular mass of around 65 kDa. It is a vital component of the NF-kB family typically found in the cytoplasm in its inactive form. Upon activation it translocates to the nucleus where it regulates the expression of target genes. NF-kB p65 is ubiquitously expressed in various cell types including immune cells and epithelial cells.
Biological function summary
NF-kB p65 plays a central role in regulating immune response and inflammation. It forms a complex with proteins such as p50 to bind DNA and modulate gene transcription. This complex is essential for cellular responses to stress and infection influencing processes such as cell proliferation and survival. It also interacts with other transcription factors and cofactors to fine-tune gene regulation.
Pathways
NF-kB p65 is integral to the canonical NF-kB signaling pathway which gets activated by signals like cytokines and bacterial products. The protein family includes several related proteins such as IκB that inhibit NF-kB activity until signal-induced degradation allows NF-kB to enter the nucleus. Another related pathway is the MAPK pathway which can interact with NF-kB signaling to coordinate cellular stress responses.
NF-kB p65 is implicated in conditions such as cancer and inflammatory diseases. Its dysregulation can lead to chronic inflammation contributing to tumor progression. Inflammatory disorders like rheumatoid arthritis also show abnormal NF-kB activity contributing to disease pathology. Additionally interactions with proteins like IKK contribute significantly to these diseases altering NF-kB signaling and gene expression patterns.