Phospho-NF-kB p65 (S536) and Total NF-kB p65 ELISA Kit
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- sELISA
Supplier Data
Sandwich ELISA - Phospho-NF-kB p65 (S536) and Total NF-kB p65 ELISA Kit (AB279874)
HeLa cells treated with/without TNF-alpha and Calyculin A.
HeLa cells were untreated or treated with TNF alpha and 100nM Calyculin A for 5 minutes.
- sELISA
Supplier Data
Sandwich ELISA - Phospho-NF-kB p65 (S536) and Total NF-kB p65 ELISA Kit (AB279874)
HeLa cells treated with/without TNF-alpha and Calyculin A.
HeLa cells were untreated or treated with TNF alpha and 100nM Calyculin A for 5 minutes.
- sELISA
Supplier Data
Sandwich ELISA - Phospho-NF-kB p65 (S536) and Total NF-kB p65 ELISA Kit (AB279874)
Positive Control.
HeLa cells were treated with TNF alpha and Calyculin A at 37°C for 5 min.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
Reactivity data
Product details
Phospho-NF-kB p65 (S536) and Total NF-kB p65 ELISA Kit (ab279874) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated NF-kB p65 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-NF-kB p65 and total NF-kB p65. An anti-pan NF-kB p65 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and NF-kB p65 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho NF-kB p65 (S536) antibody is added to detect phosphorylated NF-kB p65. In the remaining wells, biotinylated anti-pan-NF-kB p65 antibody is used to detect pan NF-kB p65. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of NF-kB p65 (S536) or pan NF-kB p65 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Properties and storage information
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Appropriate long-term storage conditions
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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.
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