Kynurenic Acid ELISA Kit

Catalog No. ABIN5696690

Human Kynurenic Acid ELISA Kit, Colorimetric assay for quantification of Human Kynurenic Acid.

Quick Overview for Kynurenic Acid ELISA Kit (ABIN5696690)

Target: Kynurenic Acid (KYNA)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 7.813 pmol/mL - 500 pmol/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Cell Samples, Plasma, Serum, Tissue Lysate

Product Details Kynurenic Acid ELISA Kit

Minimum Detection Limit: 7.813 pmol/mL

Purpose: Human KYN (Kynurenine) ELISA Kit

Analytical Method: Quantitative

Specificity: Specifically binds with KYN , no obvious cross reaction with other analogues.

Sensitivity: 4.688 pmol/mL

Components:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Lyophilized Biotin-labeled Antibody(Concentrated)
• Biotin-labeled Antibody
• HRP-Streptavidin Conjugate(SABC
• 100X)
• TMB Substrate
• Sample Dilution Buffer
• Stop Solution
• Plate Sealer
• Product Description

Material not included:

• Microplate reader (wavelength: 450nm)
• 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
• Automated plate washer or multi-channel pipette/5 mL pipettor
• Precision single (0.5-10μL
• 5-50μL
• 20-200μL
• 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
• Sterile tubes and Eppendorf tubes with disposable tips
• Absorbent paper and loading slot
• Deionized or distilled water

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

Competitive ELISA, Coated with Antigen

Sample Volume: 50 μL

Assay Time: 2 h

Plate: Pre-coated

Protocol: Step 1: Wash the plate twice before adding the standard and sample. Step 2: Add 50 µL standard or sample into each well. Immediately add 50 µL biotin-labeled antibody working solution into each well(The tip can't be reused after touching the liquid.), gently shake the plate for 1 min, seal closely and statically incubate for 45 minutes at 37 °C. Washing: Wash the plate three times and immerse for 1 min each time. Step 3: Add 100 µL HRP-Streptavidin Conjugate (SABC) working solution into each well, seal the plate and statically incubate for 30 minutes at 37 °C. Washing: Wash the plate five times and immerse for 1 min each time. Step 4: Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C. (Accurate TMB visualization control is required.) Step 5: Add 50 µL stop solution. Read at 450nm immediately and calculate.

Reagent Preparation:

• Bring all reagents and samples to room temperature for 20 minutes before use.
• Wash Buffer: If crystals have formed in the concentrate, you can warm it with 40 °C water bath Concentrated Wash Buffer into 750 mL Wash Buffer with deionized or distilled water. Put unused solution back at 2-8 °C.
• Standards:
  1. Add 1 mL Sample Dilution Buffer into one Standard tube (labeled as zero tube), keep the tube at room temperature for 10 minutes and mix them thoroughly. Note: If the standard tube concentration higher than the range of the kit,please dilute it and labeled as zero tube.
  2. Label 7 EP tubes with 1/2, 1/4, 1/8, 1/16, 1/32, 1/64 and blank respectively. Add 0.3 mLof the Sample Dilution Buffer into each tube. Add 0.3 mLof the above Standard solution (from zero tube) into 1st tube and mix them thoroughly. Transfer 0.3 mL from 1st tube to 2nd tube and mix them thoroughly. Transfer 0.3 mL from 2nd tube to 3rd tube and mix them thoroughly, and so on. Sample Dilution Buffer was used for the blank control. Note: It is best to use Standard Solutions within 2 hours.
• Preparation of Biotin-labeled Antibody Working Solution:
  Prepare it within 1 hour before experiment.
  1. Dissolve: Add 70ul purified water into tube and mix them thoroughly, after the biotin-labeled antibody is dissolved, please store it at 2-8°C.
  2. Calculate required total volume of the working solution: 0.05ml/well x quantity of wells. (Allow 0.1-0.2 mLmore than the total volume.)
  3. Dilute the Biotin-detection antibody with Antibody Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1 µL Biotin-labeled antibody into 99 µL Antibody Dilution Buffer.)
• Preparation of HRP-Streptavidin Conjugate (SABC) Working Solution:
  Prepare it within 30 minutes before experiment.
  1. Calculate required total volume of the working solution: 0.1ml/well x quantity of wells. (Allow 0.1-0.2 mLmore than the total volume.)
  2. Dilute the SABC with SABC Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1 µL of SABC into 99 µL of SABC Dilution Buffer.)

Restrictions: For Research Use only

Handling

Handling Advice: Avoid direct light exposure to antibody solutions

Storage: 4 °C

Storage Comment: Store sealed Kit at 2-8° C. Do Not Cryopreserve. Store ELISA Microplate for 1 month at 2-8°C, Store for 12 month at -20°C. Put the leftover strips into a sealed foil bag with the desiccant

Expiry Date: 12 months

Target Details for Kynurenic Acid

Target: Kynurenic Acid (KYNA)

Alternative Name: KYN (Kynurenine)

Target Type: Chemical

Background: Kynurenine, KYN