IL-1 beta ELISA Kit

Produktnummer ABIN7671658

Das Human IL-1 beta ELISA Kit (ABIN7671658) ist ein Colorimetric ELISA Kit zur Detektion von Human IL-1 beta.

Kurzübersicht für IL-1 beta ELISA Kit (ABIN7671658)

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 3.906 pg/mL - 250 pg/mL

Applikation: ELISA

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

Güteklasse: Quick

Produktdetails IL-1 beta ELISA Kit

Untere Nachweisgrenze: 3.906 pg/mL

Verwendungszweck: Human IL-1β (Interleukin 1 Beta) QuickTest ELISA Kit

Analytische Methode: Quantitative

Spezifität: Specifically binds with IL-1β , 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 0-6.5pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=16) 1/2 dilution ND-7.2pg/mL Human peripheral blood lymphocyte were cultured with 10 %FBS + 1640 + double antibody, and the cell culture supernatant was detected after 12 hours. undiluted ND Human peripheral blood lymphocyte were cultured with 10 %FBS + 1640 + double antibody+1 μg/mL PHA, and the cell culture supernatant was detected after 12 hours. 1/10-1/20 dilution 4011 pg/mL THP-1 cells were cultured with 5 %FBS + 1640 + double antibody+1 μg/mL LPS, and the cell culture supernatant was detected after 12 hours. 1/5 dilution 996 pg/mL 2. KO sample validation (Detect IL-1β 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 100 ng/mL LPS for 6 hours. The cell culture supernatant was taken for detection. 1/10 dilution 1356 pg/mL KO THP-1 cells were treated with 80nM TPA and cultured overnight, and then stimulated with 100 ng/mL LPS for 6 hours. The cell culture supernatant was taken for detection. undiluted ND KO THP-1 cells were treated with 80nM TPA and cultured overnight, and then stimulated with 100 ng/mL LPS for 6 hours. After that, 300 ng/mL Brefeldin A (BFA) was added and cultured for 3 hours. Add cell lysis buffer(Catalogue No.:E050), collect the lysate solution (total protein concentration measured by BCA assay: 1.66 mg/mL) to detect. undiluted ND Note:ND is lower than the sensitivity of the kit and was not detected 3. Capture antibody and detection antibody by WB KO validation (Detect IL-1β KO THP-1 cells):

Sensitivität: 2.344 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-1 beta

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Andere Bezeichnung: IL-1beta

Hintergrund: Interleukin-1 beta, IL-1 beta, Catabolin, IL1B, IL1F2

UniProt: P01584

Pathways: NF-kappaB Signalweg, Interferon-gamma Pathway, TLR Signalweg, Negative Regulation of Hormone Secretion, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Myometrial Relaxation and Contraction, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Autophagie, Cancer Immune Checkpoints, Inflammasome