IL-10 ELISA Kit

Produktnummer ABIN7671665

Human IL-10 ELISA Kit, Colorimetric Assay für die Quantifizierung von Human IL-10.

Kurzübersicht für IL-10 ELISA Kit (ABIN7671665)

Target: IL-10 (IL10) (Interleukin 10 (IL10))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 7.813 pg/mL - 500 pg/mL

Applikation: ELISA

Proben: Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate

Güteklasse: Quick

Produktdetails IL-10 ELISA Kit

Untere Nachweisgrenze: 7.813 pg/mL

Verwendungszweck: Human IL-10 (Interleukin 10) QuickTest ELISA Kit

Analytische Methode: Quantitative

Spezifität: Specifically binds with IL-10 , 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. (When the sample content 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=18) undiluted ND-10.7pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=20) undiluted ND-11.3pg/mL Human peripheral blood cells were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 10 μg/mL PHA for 24 hours. The cell culture supernatants were assayed for levels of human IL-10. undiluted 118pg/mL THP-1 cells were treated with 80nM TPA and cultured overnight.The cell culture supernatants were assayed for levels of human IL-10. undiluted ND THP-1 cells were treated with 80nM TPA and cultured overnight. Then, stimulate with 100 ng/mL LPS for 6 hours and take the cell culture supernatant were assayed for levels of human IL-10. undiluted 408pg/mL 2. KO sample validation (Detect IL-10 KO THP-1 cells): Sample Type Dilution Ratio Content Wild THP-1 cells were treated with 80nM TPA and cultured overnight, and then stimulated with 200 ng/mL LPS for 6 hours. The cell culture supernatants were assayed for levels of human IL-10. 1/5 dilution 506pg/mL KO THP-1 cells were treated with 80nM TPA and cultured overnight, and then stimulated with 200 ng/mL LPS for 6 hours. The cell culture supernatants were assayed for levels of human IL-10. undiluted ND 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-10 protein Ser19-Asn178B. E. coli-derived Human IL-10 protein Ser19-Asn178

Sensitivität: 4.688 pg/mL

Benötigtes Material:

• 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

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Kommentare:

Sandwich ELISA, Double Antibody

Probenmenge: 50 μL

Testdauer: 2 h

Plattentyp: Pre-coated

Protokoll: Step 1: Take out the required plate wells, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37 °C. Washing: Wash the plate twice without immersion. Step 2: Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37 °C. Washing: Wash the plate five times without immersion. Step 3: Add 90 µL TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37 °C. (Accurate TMB visualization control is required.) Step 4: Add 50 µL stop solution. Read at 450nm immediately and calculate.

Testdurchführung:

1. Take out the required strips, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into relevant well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure complete mixing then statically incubate for 60 minutes at 37 °C.
2. Wash the plate twice without immersion.
3. Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
4. Wash the plate five times without immersion.
5. 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.).
6. Add 50 µL stop solution. Read at 450nm immediately and calculate.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid direct light exposure to antibody solutions

Lagerung: 4 °C

Informationen zur Lagerung: 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

Haltbarkeit: 12 months

Details zu IL-10

Target: IL-10 (IL10) (Interleukin 10 (IL10))

Andere Bezeichnung: IL-10

Hintergrund: Interleukin-10, IL-10, Cytokine synthesis inhibitory factor, CSIF, IL10

UniProt: P22301

Pathways: Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Production of Molecular Mediator of Immune Response, Maintenance of Protein Location, Cancer Immune Checkpoints