Phospho-Hsp27 (S82) and Total Hsp27 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Hsp27 (S82) and Total Hsp27 in Human in Cell Lysate samples.
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Phospho-Hsp27 (S82) and Total Hsp27 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Hsp27 (S82) and Total Hsp27 in Human in Cell Lysate samples.
Phospho-Hsp27 (S82) and Total Hsp27 ELISA Kit (ab279823) 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 Hsp27 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-Hsp27 and total Hsp27. An anti-pan Hsp27 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Hsp27 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho Hsp27 (S82) antibody is added to detect phosphorylated Hsp27. In the remaining wells, biotinylated anti-pan-Hsp27 antibody is used to detect pan Hsp27. 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 Hsp27 (S82) or pan Hsp27 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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Hsp27 also known as HSPB1 is a small heat shock protein with a molecular weight of approximately 27 kilodaltons. This protein is expressed in various tissues including muscle heart and brain. It functions as a molecular chaperone that stabilizes unfolded proteins preventing their aggregation. Hsp27 undergoes phosphorylation at specific residues which modulates its chaperone activity and interaction with other proteins.
Hsp27 plays a critical role in cellular stress response by regulating actin cytoskeleton dynamics and inhibiting apoptosis. It forms part of a complex that includes other proteins such as alphaB-crystallin. This complex facilitates the reorganization of proteins under stress conditions enhancing cell survival during oxidative stress or thermal shock. Hsp27 also modulates inflammatory responses and has been shown to affect cell migration.
Hsp27 integrates into the apoptosis and inflammation pathways. It interacts with apoptotic machinery such as caspase proteins to protect cells by hindering apoptosome formation. Additionally Hsp27 can engage with pathways involving the nuclear factor-kappa B (NF-kB) impacting inflammatory signaling. CPTC (carboxyl-pyrene-trioctylamine) can modulate these pathways by altering Hsp27 function and interactions.
Hsp27 has connections to neurodegenerative diseases and cancer. In neurodegenerative conditions such as Alzheimer's disease its chaperone activity is thought to protect neurons from misfolded protein aggregates. In cancer Hsp27 supports tumor cell survival and resistance to chemotherapy by interacting with proteins like Akt and p53. These interactions highlight the complex role of Hsp27 in modulating cellular responses in various pathological states.
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Positive Control.
HeLa cells were treated with PMA at 37°C for 20 mins.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
HeLa cells treated/untreated with PMA. HeLa cells were untreated or treated with 250 nM PMA for 20 mins.
HeLa cells treated/untreated with PMA. HeLa cells were untreated or treated with 250 nM PMA for 20 mins.
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