IL-33 ELISA Kit

Produktnummer ABIN7671718

Das Human IL-33 ELISA Kit (ABIN7671718) ist ein Colorimetric ELISA Kit zur Detektion von Human IL-33.

Kurzübersicht für IL-33 ELISA Kit (ABIN7671718)

Target: IL-33 (IL33) (Interleukin 33 (IL33))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 3.125 pg/mL - 200 pg/mL

Applikation: ELISA

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

Güteklasse: Quick

Produktdetails IL-33 ELISA Kit

Untere Nachweisgrenze: 3.125 pg/mL

Verwendungszweck: Human IL-33 (Interleukin-33) QuickTest ELISA Kit

Analytische Methode: Quantitative

Spezifität: Specifically binds with IL-33 , 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=20) undiluted ND-21pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=22) undiluted ND-24pg/mL DLD-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 1 μg/mL LPS stimulation for 12 hours. Add cell lysis buffer(Catalogue No.:E050), the cell lysate solution (total protein concentration measured by BCA assay: 2.8 mg/mL) were assayed for levels of human IL-33. undiluted 19pg/mg 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-33. 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, incubated overnight. The cells were then stimulated with 1 μg/mL LPS stimulation for 12 hours, and cell culture supernatants were assayed for levels of human IL-33. 1/2 dilution 183pg/mL 2. KD sample validation (Detect IL-33-KD THP-1 cells): Sample Type Dilution Ratio Content Wild 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, incubated overnight. The cells were then stimulated with 1 μg/mL LPS stimulation for 12 hours, and cell culture supernatants were assayed for levels of human IL-33. 1/2 dilution 213pg/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, incubated overnight. The cells were then stimulated with 1 μg/mL LPS stimulation for 12 hours, and cell culture supernatants were assayed for levels of human IL-33. undiluted 26pg/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. E. coli-derived Human IL-33 protein Ser112-Thr270

Sensitivität: 1.875 pg/mL

Bestandteile:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Cap/Det Ab (Ready to use
• blue)
• TMB Substrate
• Sample Dilution Buffer
• Stop Solution
• Plate Sealer
• Product Description

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-33

Target: IL-33 (IL33) (Interleukin 33 (IL33))

Andere Bezeichnung: IL-33

Hintergrund: Interleukin-33, IL-33, Interleukin-1 family member 11, IL-1F11, Nuclear factor from high endothelial venules, NF-HEV, IL33, C9orf26, IL1F11, NFHEV

UniProt: O95760

Pathways: Production of Molecular Mediator of Immune Response