Hexanoyl-Lys adduct ELISA Assay

$1,260.00

The Hexanoyl-Lys adduct (HEL) ELISA assay kit is intended for the quantitative determination of Hexanoyl­Lysine adduct in urine, serum or biological samples by enzyme linked immunoassay (ELISA). The Hexanoyl­Lysine adduct ELISA assay kit is for research use only and not to be used in diagnostic procedures.

SKU: HEL39-K01 Categories: , , Tags: ,

Hexanoyl-Lys adduct ELISA Assay

The Hexanoyl-Lys adduct ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 2.6 nmol/L
Dynamic Range: 2.6 – 264 nmol/L
Incubation Time: Overnight
Sample Type:Serum, Urine, Biological Fluids
Sample Size: 50 µL
Alternative Names: HEL, Hexanoyl­Lysine adduct


Sample Preparation
Urine sample: Dilution of samples at least 4 times using phosphate buffered saline (PBS at pH 7.4). For urine sample from experimental animal such as dogs and cats, 10 times or 20 times dilution is recommended. If insoluble materials are observed, remove them by centrifugation. If the urine contains proteins, treat the urine by the same procedure as for serum sample.
Serum sample: Prepare “Enzyme reagent”, by dissolving 14 mg/mL of alpha-chymotrypsin in PBS (pH7.4). Dilute the serum sample at least 2 times using PBS (pH7.4). Mix 300 µL of diluted sample and 60 µL of “Enzyme reagent”. Incubate at 37 °C overnight. Filtrate using ultra filter with cut-off molecular weight 10kDa (for example Microcon YM-10, Millipore), and remove enzymes. Apply the filtrate to ELISA.
*This is an example of procedure. Please investigate optimum condition depending on the sample.


Assay Background

Hexanoyl-Lys adduct (HEL) is formed by the reaction of linoleic acid hydroperoxide and Lysine, and a biomarker for oxidative stress. The Hexanoyl­Lysine adduct assay kit is a competitive enzyme-linked immunosorbent assay for quantitative measurement of hexanoyl-Lys adduct. This kit is based on the monoclonal antibody clone 5H4, which is specific for HEL. Suitable for urine, serum and other biological samples.


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Additional Information

Assay Procedure


  1. Prepare Washing solution by mixing one bottle of Washing buffer(x5) and 100 mL of distilled water.
  2. Add 50 µL of Standards(A-F) or sample per well. For the Blank well, add 100 µL of Washing solution.
  3. Add 50 µL of Primary Antibody to all wells except Blank well. Seal the microtiter plate tightly with Plate Seal. Mix gently by shaking the microtiter plate horizontally. Incubate at 4°C overnight.
  4. Reconstitute Secondary Antibody with one bottle of Secondary Antibody Buffer. This is stable for 1 week at 4°C.
  5. Remove the plate seal, and pour off the contents of microtiter plate by turning the plate upside down. The use of aspirator is not recommended. Remove the remaining solution by blotting the plate against clean paper towel. Add 250 µL of Washing solution to each well, mix gently by horizontal shaking, and remove the contents similarly. Repeat washing procedure twice and remove the remaining solution of the well.
  6. Add 100 µL of Secondary Antibody to all wells. Seal the microtiter plate tightly with Plate Seal. Mix gently by shaking the microtiter plate horizontally. Incubate at room temperature for 1 hour.
  7. Prepare Chromogen solution. Add 120 µL of Chromogen to Chromogen. Buffer bottle. Please note that Chromogen solution should be prepared just before use. Alternatively, dilute Chromogen with 100 volumes of Chromogen Buffer.
  8. Remove the plate seal, and wash the plate as mentioned at STEP F for 3 times. Remove the remaining solution of the well.
  9. Add 100 µL of Chromogen solution to all well, and incubate at room temperature for 15 minutes in the dark.
  10. Add 100 µL of Stop Solution to all wells, mix gently, wait for 3 minutes, and measure the absorbance at 450 nm.

Manual

Product Manual