Leukotriene C4 ELISA Kit

Sample

Tissue culture supernatants

Detection Method

cELISA

Intended Use

LTC₄ ELISA Kit is a competitive assay that can be used for quantification of LTC₄ in culture media and other sample matrices. The assay has a range from 17.1-5,000 pg/ml, an average sensitivity(80% B/B₀) of 21.2 pg/ml, and a lower limit of detection (LLOD) of 7.4 pg/ml.

Contents of Kit

1. Leukotriene C₄ ELISA Antiserum, 1 vial/100 dtn
2. Leukotriene C₄ AChE Tracer, 1 vial/100 dtn
3. Leukotriene C₄ ELISA Standard, 1 vial
4. ELISA Buffer Concentrate(10×), 2 vials/10 ml
5. Wash Buffer Concentrate(400×), 1 vial/5ml
6. Polysorbate 20, 1 vial/3 ml
7. Mouse Anti-Rabbit lgG Coated Plate, 1 plate, 96-Well
8. Cover Sheet, 1 cover
9. Ellman's Reagent, 3 vials/100 dtn
10. ELISA Tracer Dye, 1 vial
11. ELISA Antiserum Dye, 1 vial

Storage

This kit wil perform as specified if stored as directed at -80°C and used before the expiration date indicated on the outside of the box.

Precision

Tissue culture samples containing a high, medium, or low level of LTC₄ were measured 60 times each using a single set of reagents.The calculated %CV is reported as intra-assay variance. A separate series of samples containing ahigh,medium,or low level of LTC₄ were measured four times each using eight independent sets of reagents.The calculated %CV is reported as inter-assay variance.

Detection Range

17.1-5,000 pg/ml

Detection Limit

Lower limit of detection (LLOD): 7.4 pg/ml.

Sensitivity

An average sensitivity(80% B/B₀): 21.2 pg/ml

General Description

Leukotriene C₄ (LTC₄) is a potent inflammatory mediator formed via the 5-lipoxygenase (5-LO) pathway in leukocytes and other 5-LO-expressing cells, including mast cells, eosinophils, and macrophages. Upon cellular stimulation or injury, 5-LO converts arachidonic acid into LTA₄ with 5(S)-HpETE as an intermediate. The subsequent conjugation of glutathione to LTA₄ by LTC₄ synthase results in LTC₄ formation. LTC₄ is rapidly metabolized by Y-glutamyl transpeptidases to LTD₄ which is then converted to LTE₄ by dipeptidases. LTC₄ is also produced via transcellular metabolism in platelets, which lack 5-LO but express LTC₄ synthase, and in endothelial and smooth muscle cells by microsomal glutathione S-transferase 2 (MGST2). LTC₄ acts on cysteinyl leukotriene 1 (CysLT₁ ) and CysLT₂ receptors to induce bronchoconstriction and vasoconstriction and is recognized as a paracrine mediator pertinent to asthma and allergic diseases.

Standard Curve