Product name15(S)-HETE ELISA Kit
Intra-assay Sample n Mean SD CV% Low conc 16 181pg/ml = 15.8% Medium conc 16 853pg/ml = 9.8% High conc 16 3929pg/ml = 3.4% Inter-assay Sample n Mean SD CV% Low conc 8 192pg/ml = 19.1% Medium conc 8 730pg/ml = 8.9% High conc 8 4335pg/ml = 6.1%
Sample typeCell culture supernatant, Urine, Serum, Plasma
Sensitivity= 69.21 pg/ml
Range78.1 pg/ml - 20000 pg/ml
Sample specific recovery Sample type Average % Range Urine = 89.6 % - % Serum = 95.9 % - % Tissue Culture Media = 100.3 % - % EDTA Plasma = 105.4 % - %
Assay durationMultiple steps standard assay
Abcam’s 15(S)-HETE in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of 15-hydroxyeicosatetraenoic acid in plasma, serum, urine, tissue culture media and other biological fluids.
A goat anti-rabbit IgG antibody has been precoated onto 96-well plates. Standards or test samples are added to the wells, along with an alkaline phosphatase (AP) conjugated-15(S)-HETE antigen and a polyclonal rabbit antibody specific to 15(S)-HETE. After incubation the excess reagents are washed away. pNpp substrate is added and after a short incubation the enzyme reaction is stopped and the yellow color generated is read at 405 nm. The intensity of the yellow coloration is inversely proportional to the amount of 15(S)-HETE captured in the plate.
15(S)-HETE (15-hydroxyeicosatetraenoic acid) is the major hydroxy derivative of arachidonic acid when acted upon by 15-lipoxygenase (15-LOX). It is also the primary monohydroxy acid synthesized by the lipoxygenase activity of Cyclooxygenase-1. Aspirin-mediated acetylation of the COX-1 enzyme results in 15(R)-HETE. Blood platelets, peripheral leukocytes, vascular smooth muscle and other cell types produce Type-1 15-LOX while prostate, lung, skin and cornea tissues produce Type-2. 15-HETE has been proposed to act as a paracrine regulator of smooth muscle and lung neutrophil recruitment due in part to its incorporation into tracheal epithelium at the sn-2 position of phosphatidylinositol. The phosphoinositol modification in turn is thought to affect signal transduction and the regulation of intracellular calcium. Interleukin-4 has been shown to regulate 15(S)-HETE expression and incorporation into cellular phospholipids. 15(S)-HETE binds to actin and the alpha-subunit of mitochondrial ATP synthase suggesting a more direct method in regulating some physiological activities. Increased levels of 15(S)-HETE are associated with asthma, rhinitis, chronic paranasal sinusitis and rheumatoid arthritis.
Compound % Cross Reactivity 15(S)-HETE 100 5,15-diHETE 1
1 13(S)-HODE 0.6 5-HETE 0.1 PGB2 0.1 PGD2 0.1 PGF2α 0.1 12(S)-HETE <0.05 12(R)-HETE <0.05 Arachidonic Acid <0.05 PGE2 <0.05 Linoleic Acid <0.05
Tested applicationsSuitable for: Competitive ELISAmore details
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 15(S)-HETE Alkaline Phosphatase Conjugate 1 x 5ml 15(S)-HETE Antibody 1 x 5ml 15(S)-HETE Standard 1 x 500µl 20X Wash Buffer Concentrate 1 x 27ml Assay Buffer 1 x 27ml Goat anti-rabbit IgG Microplate (12 x 8 wells) 1 unit Plate Sealer 2 units pNpp Substrate 1 x 20ml Stop Solution 1 x 5ml
- 15(S)-hydroperoxy tetraenoic eicosatetraenoic acid
Our Abpromise guarantee covers the use of ab133035 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Competitive ELISA||Use at an assay dependent concentration.|
This product has been referenced in:
- Sandstedt M et al. Hypoxic cardiac fibroblasts from failing human hearts decrease cardiomyocyte beating frequency in an ALOX15 dependent manner. PLoS One 13:e0202693 (2018). Read more (PubMed: 30138423) »
- Vijil C et al. Arachidonate 15-lipoxygenase enzyme products increase platelet aggregation and thrombin generation. PLoS One 9:e88546 (2014). Read more (PubMed: 24533104) »