9 (±) Hydroxyoctadecadienoic (HODE) Acid ELISA Assay Kit

$410.00

The Eagle Biosciences 9 (±) Hydroxyoctadecadienoic (HODE) Acid ELISA Assay kit is for the quantitative determination of Hydroxyoctadecadienoic (HODE) Acid in biological fluids by a microplate enzyme immunoassay (ELISA).

SKU: 9HY39-K01 Categories: , ,

9 (±) Hydroxyoctadecadienoic (HODE) Acid ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.1 ng/ml
Dynamic Range: 0.1 – 500 ng/ml
Incubation Time: 3 hour
Sample Type: Biological Fluids
Sample Size: 1 mL

Product manufactured in the USA

Additional Information

Assay Background

Linoleic acid, the predominant polyunsaturated fatty acid (PUFA) in the human diet, can be metabolized by cyclooxygenase, lipoxygenase and P450 enzymes.  The hydroxyoctadecadienoic acid (HODE) derivatives of linoleic acid, 9(R)-HODE, 9(S)-HODE and 13(S)-HODE, are the most widely distributed of the known linoleic acid metabolites. These compounds exhibit interesting biologic activities, including the regulation of platelet function, maintenance of vascular thromboresistance and transduction of the cellular responses to certain growth factors. HODE derivatives may also influence certain pathological states including psoriasis, the development of atherosclerosis and the development of cancer.  This assay measures the level of total 9-HODE, which includes both 9(S)-HODE and 9(R)-HODE, in biological samples.

Assay Principle

9 (±) Hydroxyoctadecadienoic (HODE) Acid ELISA Assay kit is a competitive immunoassay (ELISA).  Briefly, the 9-HODE present in the samples or standards competes with 9(±)-HODE conjugated to horseradish peroxidase [9(±)-HODE-HRP] for binding to an antibody specific for 9(±)-HODE that is precoated on a microplate.  The peroxidase activity of 9(±)-HODE-HRP results in color development when a substrate is added.  The intensity of the color is proportional to the amount of 9(±)-HODE-HRP bound and is inversely proportional to the amount of unconjugated 9-HODE present in the samples or standards.

  1. Add 100 µL of Standards or Samples to the corresponding wells on the microplate in duplicate.  See Scheme I for a sample plate layout.
  2. Add 100 µL of diluted 9(±)-HODE-HRP Conjugate to each well. Incubate at room temperature for two hours.
  3. Wash the plate three times with 300 µL of diluted Wash Buffer per well.
  4. Add 200 µL of TMB Substrate to each well. Incubate at room temperature for 45-60 minutes.
  5. Add 50 µL of 3 N H2SO4 to each well to stop the reaction.
  6. Read the plate at 450 nm.

Cross Reactivity Data

9(±)-HODE 100.0%
9(S)-HODE 100.0%
9(R)-HODE 100.0%
13(S)-Hydroxyoctadecadienoic Acid 1.2%
13(R)-Hydroxyoctadecadienoic Acid 1.2%
9-oxo-Octadecadienoic Acid 1.2%
13-oxo-Octadecadienoic Acid 2.4%
11(S)-HETE 0.0%
15(S)-HETE 0.0%

Manual

Product Manual