12(S)-HETE ELISA Kit (ab133034)
Key features and details
- Sensitivity: 146.3 pg/ml
- Range: 195 pg/ml - 50000 pg/ml
- Sample type: Cell culture supernatant, Plasma
- Detection method: Colorimetric
- Assay type: Competitive
Overview
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Product name
12(S)-HETE ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Buffer 16 342pg/ml = 5.2% Buffer 16 1153pg/ml = 10.1% Buffer 16 4762pg/ml = 15.5% Inter-assay Sample n Mean SD CV% Buffer 224pg/ml = 4.1% Buffer 1127pg/ml = 9.1% Buffer 5294pg/ml = 20.8% -
Sample type
Cell culture supernatant, Plasma -
Assay type
Competitive -
Sensitivity
= 146.3 pg/ml -
Range
195 pg/ml - 50000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture media = 94 % - % Hep Plasma = 104 % - % EDTA Plasma = 97 % - % -
Assay duration
Multiple steps standard assay -
Product overview
Abcam’s 12(S)-HETE in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of 12(S)-HETE in cell culture supernatants and plasma (heparin, EDTA).
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-12(S)-HETE antigen and a polyclonal rabbit antibody specific to 12(S)-HETE. After incubation the excess reagents are washed away. pNpp substrate is added and after a short incubation the alkaline phosphatase 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 12(S)-HETE captured in the plate.
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Notes
12(S)-HETE is the stereo specific hydroxy product from the reduction of 12(S)-hydroperoxy tetraenoic eicosatetraenoic acid [12(S)-HpETE], which itself is a 12-lipoxygenase metabolite of arachidonic acid. 12(S)-HETE has been shown to be chemotactic and chemokinetic for polymorphonuclear leukocytes and vascular smooth cells. It also acts as a second messenger in angiotensin-II induced aldosterone production. Evidence also suggests that 12(S)-HETE is involved in suppressing renin production, stimulating insulin secretion by pancreatic tissue, inducing endothelial cell retraction and tumor cell adhesion.
Cross Reactivity
Compound % Cross Reactivity 12(S)-HETE 100 12(R)-HETE 2.5 15-HETE 0.3 5(S)-HETE 0.2 8,15-diHETE 0.1 5,15-diHETE 0.1 PGE2 0.1 PGF2α 0.1 PGD2 0.1 6-keto-PGF1α 0.1 Thromboxane B2 0.1 Arachidonic Acid 0.1 Leukotriene B4 0.1 Leukotriene C4 0.1 Leukotriene D4 0.1 Leukotriene E4 0.1 8-HETE 9-HETE 11-HETE -
Platform
Microplate
Properties
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Storage instructions
Please refer to protocols. -
Components 1 x 96 tests 12(S)-HETE Antibody 1 x 5ml 12(S)-HETE Conjugate 1 x 5ml 12(S)-HETE Standard 1 x 0.5ml 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 -
Research areas
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Relevance
12(S)-HETE is the stereo specific hydroxy product from the reduction of 12(S)-hydroperoxy tetraenoic eicosatetraenoic acid [12(S)-HpETE], which itself is a 12-lipoxygenase metabolite of arachidonic acid. 12(S)-HETE has been shown to be chemotactic and chemokinetic for polymorphonuclear leukocytes and vascular smooth cells. It also acts as a second messenger in angiotensin-II induced aldosterone production. Evidence also suggests that 12(S)-HETE is involved in suppressing renin production, stimulating insulin secretion by pancreatic tissue, inducing endothelial cell retraction and tumor cell adhesion.