IL-13 ELISA Kit (Interleukin 13)

Details for Product IL13 ELISA Kit No. ABIN1979388
Target
Reactivity
Pig
Alternatives
Kits with alternative reactivity to:
11
6
6
6
6
5
4
4
4
3
3
3
2
2
2
2
Detection Method
Colorimetric
Method Type
Sandwich ELISA
Detection Range
16-15000 pg/mL
Minimum Detection Limit
16 pg/mL
Application
ELISA
Options
Purpose Porcine IL-13 ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Serum, Plasma, Cell Culture Supernatant
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with the following cytokines tested: porcine CCL3L1, IFN alpha, IL-1 alpha, IL-1R alpha, IL-17A, IL-18, MIG, MIP-1 beta, PECAM-1.
Cross-Reactivity (Details) This ELISA kit shows no cross-reactivity with the following cytokines tested: porcine CCL3L1, IFN alpha, IL-1 alpha, IL-1R alpha, IL-17A, IL-18, MIG, MIP-1 beta, PECAM-1.
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Target
Alternative Name IL-13 (IL13 ELISA Kit Abstract)
Gene ID 396721
UniProt Q95J68
Pathways JAK-STAT Signaling, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Proton Transport
Application Notes Recommended Dilution for serum and plasma samples5 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. 2. Sample dilution: If your samples need to be diluted, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 5 fold. The Porcine IL-13 ELISA Kit Protocol 3 Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator. 3. Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use. 4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent (Item E) into Item C vial to prepare a 50 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 150 µL IL-13 standard solution from the vial of Item C, into a tube with 350 µL 1x Assay Diluent to prepare a 15,000 pg/ml standard solution. Pipette 400myl 1x Assay Diluent into each tube. Use the 15,000 pg/ml standard solution to produce a Dilution series (shown below). Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/ml). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 150 µL standard + 350 µL 15,000 5,000 1,667 555.6 185.2 61.73 20.58 0 pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to yield 400 mL of 1x Wash Buffer. The Porcine IL-13 ELISA Kit Protocol 4 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 800-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 15 µL of HRP-Streptavidin concentrate into a tube with 12 mL 1x Assay Diluent to prepare a 800-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Assay Procedure
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking. 3. Discard 4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Assay Precision Intra-Assay: CV<10%
Inter-Assay: CV<12%
Restrictions For Research Use only
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months
Images
Image no. 1 for Interleukin 13 (IL13) ELISA Kit (ABIN1979388) Interleukin 13 (IL13) ELISA Kit