IL27 ELISA Kit

Catalog No. ABIN5522128

This Colorimetric ELISA kit is designed for the quantitative measurement of Human IL27.

Quick Overview for IL27 ELISA Kit (ABIN5522128)

Target: IL27 (Interleukin-27 Subunit alpha (IL27))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.156 ng/mL - 10 ng/mL

Application: ELISA

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

Product Details IL27 ELISA Kit

Minimum Detection Limit: 0.156 ng/mL

Purpose: Human IL-27 (Interleukin-27 subunit alpha) ELISA Kit

Analytical Method: Quantitative

Specificity: Specifically binds with IL-27 , no obvious cross reaction with other analogues. Recommended Sample Dilution Ratio The following table shows the recommended dilution ratios for this kit for a limited number of samples for your reference only. (The matrix components in serum/plasma will affect the test results, which it need to be diluted at least 1/2 with Sample Dilution Buffer before testing! When the content of other samples is very low, the original solution can be added without dilution, but it is necessary to ensure that the pH is between 6.8 and 8.0, and it does not contain more than 10 % organic solvents or high-concentration protein denaturants.) 1. Common sample validation: Sample Type Recommended Dilution Ratio Content Healthy serum (n=19) 1/2 dilution 200-1250pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=20) 1/2 dilution 190-1290pg/mL THP-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate and incubated overnight. The cell culture supernatants were assayed for levels of human IL-27. undiluted ND THP-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate and incubated overnight. The cells were then stimulated with 100 ng/mL LPS for 12 hours, and cell culture supernatants were assayed for levels of human IL-27. undiluted 956pg/mL 2. KD sample validation (Detect IL-27-KD THP-1 cells): Sample Type Dilution Ratio Content Wild THP-1 cells THP-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate and incubated overnight. The cells were then stimulated with 100 ng/mL LPS for 12 hours, and cell culture supernatants were assayed for levels of human IL-27. undiluted 1062pg/mL THP-1 cells (+siRNA) were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate and incubated overnight. The cells were then stimulated with 100 ng/mL LPS for 12 hours, and cell culture supernatants were assayed for levels of human IL-27. undiluted 174pg/mL Note:ND is lower than the sensitivity of the kit and was not detected 3. Verified recombinant protein: (It is a normal phenomenon that some proteins have weak detection signals or cannot be detected at all due to differences in tags, sequences or protein activities) A. HEK293-derived Human IL-27 protein Phe29-Pro243(IL-27A)B. CHO-derived Human IL-27 protein Phe29-Pro243(IL-27A)

Sensitivity: 0.094 ng/mL

Components:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• 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:

Sandwich ELISA, Double Antibody

Sample Volume: 100 μL

Assay Time: 4 h

Plate: Pre-coated

Protocol: Step 1: Add 100 µL standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37 °C. Washing: Wash the plate twice without immersing. Step 2: Add 100 µL biotin-antibody working solution, seal the plate and statically incubate for 60 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, 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.

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 IL27

Target: IL27 (Interleukin-27 Subunit alpha (IL27))

Alternative Name: Interleukin-27 subunit alpha

Background: Interleukin-27 subunit alpha, IL-27 subunit alpha, IL-27-A, IL27-A, Interleukin-30, p28, IL27, IL27A, IL30

UniProt: Q8NEV9