This is an ELISA (Enzyme-Linked ImmunoSorbent Assay) for the quantitative analysis of Leukotriene C4/D4/E4 levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and Leukotriene C4/D4/E4 in the sample for a limited number of binding sites.
Contents of Kit
1. EIA BUFFER: 30 mL. Provided to dilute enzyme conjugate and Leukotriene C4 standards. NOTE: PLEASE ONLY USE THE EIA BUFFER THAT IS PROVIDED WITH THIS KIT LOT.
2. WASH BUFFER (10X): 20 mL. Dilute 10-fold with deionized water. Diluted wash buffer is used to wash all unbound enzyme conjugate, samples and standards from the plate after the one hour incubation.
3. K-BLUE SUBSTRATE: 20 mL. Stabilized 3,3', 5,5' Tetramethylbenzidine (TMB) plus Hydrogen Peroxide (H2O2) in a single bottle. It is used to develop the color in the wells after they have been washed. Keep refrigerated. LIGHT SENSITIVE.
4. EXTRACTION BUFFER (5X): 30mL. Dilute 5-fold with deionized water. This is used for diluting extracted and non-extracted samples.
5. LEUKOTRIENE C4/D4/E4 ENZYME LYOPHILIZED CONJUGATE: Two vials of lyophilized LTC4/D4/E4 horseradish peroxidase conjugate. Reconstitution with 75 µL of deionized water results in a 50:1 concentrate. Store lyophilized vials at -20°C. Store reconstituted vials at 4°C. Blue capped vials.
6. LEUKOTRIENE C4 STANDARD: 100 µL. Leukotriene C4 standard at the concentration of 1 µg/mL. Green capped vial.
7. LEUKOTRIENE C4/D4/E4 ANTIBODY-COATED MICROPLATE: A 96 well Maxisorp™ Nunc microplate with anti-LTC4/D4/E4 monoclonal rat antibody precoated on each well. The plate is ready to use as is. DO NOT WASH!
Lyophilized conjugate: -20°C or less
Do not freeze kit reconstituted conjugate
All other kit components: 4°C
Leukotriene C4/D4/E4(LTC4/D4/E4) are potent mediators of immediate hypersensitivity, bronchoconstriction, smooth muscle contraction, increased vascular permeability and epithelial mucous secretion. They are derived from arachidonic acid through the 5-lipoxygenase pathway. The monoclonal antisera in this kit recognizes LTC4, LTD4 and LTE4 for a combined quantitative value.