Quinolinic Acid ELISA Assay
Quinolinic Acid ELISA Assay Developed and Manufactured in France by ImmuSmol
Size: 1×96 wells
Sensitivity: < 6 ng/mL
Standard Range: 25 – 2000 ng/mL
Incubation Time: Overnight
Sample Type: Serum
Sample Size: 25µL
Alternative Names: QUIN Acid ELISA, QA ELISA
For Research Use Only
Specificity: No significant cross-reactivity was observed with Quinolinic Acid analogs such as Kynurenic acid, Xanthurenic acid, Kynurenine, Picolinic acid, 3Hydroxy-Anthranilic Acid
Sample Collection and Storage: Serum: Do not use lipemic, haemolytic samples, as well as samples containing precipitates or fibrin strands. Store samples at 2-8°C for up to 48h or -20°C for longer period (up to 6 months).
After extraction and derivatization Quinolinic acid is quantitatively determined by ELISA. The competitive Quinolinic Acid ELISA uses the microtiter plate format. The antigen is bound to the solid phase of the microtiter plate. The processed standards, controls and samples and the solid phase bound analyte compete for a fixed number of antiserum binding sites. When the system is in equilibrium, free antigen and free antigen-antiserum complexes are removed by washing. The antibody bound to the solid phase is detected by an anti-rabbit IgG-peroxidase conjugate using TMB as a substrate. The reaction is monitored at 450 nm. Quantification of unknown samples is achieved by comparing their absorbance with a reference curve prepared with known standards.
Products Related to Quinolinic Acid ELISA Assay
L-Quinolinic Acid (Urine) ELISA Assay Kit
Quinolinic Acid Polyclonal Antibody
Quinolinic Acid Monoclonal Antibody
Allow all reagents and samples to reach room temperature and mix thoroughly by gentle inversion before use. Duplicate determinations are recommended. It is recommended to number the strips of the microwell plate before usage to avoid any mix-up.
The binding of the antisera and of the enzyme conjugate and the activity of the enzyme are temperature dependent, and the absorbance values may vary if a thermostat is not used. The higher the temperature, the higher the extinction values will be. Corresponding variations also apply to the incubation times. The optimal temperature during the Enzyme Immunoassay is between 20 – 25 °C.
- Pipette 25 µL of the standards, controls and samples into the appropriate wells of the Reaction plate.
- Add 25 µL of Reaction Diluent into all wells and mix shortly.
- Add 100 µL of the Acylation Reagent into all wells and mix shortly.
- Cover the plate with Adhesive foil and incubate 2 hours at 37°C.
- Use 40 µL for the ELISA!
- Mix shortly (2 min on a shaker at 500 rpm) and pipette 40 µL of the prepared standards, controls and samples into the appropriate wells of the Quinolinic acid Microtiter Strips.
- Pipette 50 µL of the QA Antiserum (refer to 6.1) into all wells and mix shortly.
- Cover the plate with Adhesive Foil and incubate for 15 – 20 h (overnight) at 2 – 8 °C.
- Remove the foil. Discard or aspirate the contents of the wells. Wash the plate 4 x by adding 300 µL of ELISA Wash Buffer, discarding the content and blotting dry each time by tapping the inverted plate on absorbent material.
- Pipette 100 µL of the Enzyme Conjugate into all wells.
- Incubate for 30 min at RT (20 – 25 °C) on a shaker (approx. 500 rpm).
- Discard or aspirate the contents of the wells. Wash the plate 4 x by adding 300 µL of ELISA Wash Buffer, discarding the content and blotting dry each time by tapping the inverted plate on absorbent material.
- Pipette 100 µL of the Substrate into all wells and incubate for 20 – 30 min at RT (20 – 25 °C) on a shaker (approx. 500 rpm). Avoid exposure to direct sunlight!
- Add 100 µL of the Stop Solution to each well and shake the microtiter plate to ensure a homogeneous distribution of the solution.
- Read the absorbance of the solution in the wells within 10 minutes, using a microplate reader set to 450 nm (if available a reference wavelength between 620 nm and 650 nm is recommended).
Typical Standard Curve