Interferon gamma ELISA Kit

Produktnummer ABIN7671628

Das Human Interferon gamma ELISA Kit (ABIN7671628) ist ein Colorimetric ELISA Kit zur Detektion von Human Interferon gamma.

Kurzübersicht für Interferon gamma ELISA Kit (ABIN7671628)

Target: Interferon gamma (IFNG)

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 15.625 pg/mL - 1000 pg/mL

Applikation: ELISA

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

Güteklasse: Quick

Produktdetails Interferon gamma ELISA Kit

Untere Nachweisgrenze: 15.625 pg/mL

Verwendungszweck: Human IFN-γ (Interferon gamma) QuickTest ELISA Kit

Analytische Methode: Quantitative

Spezifität: Specifically binds with IFN-γ , 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=21) undiluted ND-22pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=21) undiluted ND-16pg/mL Human peripheral blood lymphocyte were cultured with 5 %FBS + 1640 + double antibody+1 μg/mL LPS, and the cell culture supernatant was detected after 12 hours. 1/10-1/25 dilution 14.6 ng/mL Human peripheral blood lymphocyte were cultured with 5 %FBS + 1640 + double antibody+10 μg/mL PHA, and the cell culture supernatant was detected after 12 hours. undiluted 250pg/mL Human peripheral blood lymphocyte were treated with 10 μg/mL PHA for 12 hours. After that, 300 ng/mL Brefeldin A (BFA) was added and cultured for 3 hours. The cell lysate (Cat No: E050) was collected, and the total protein concentration of 1.56 mg/mL was detected by BCA. 1/2 dilution 1.32 ng/mg(total protein) 2. KO sample validation (Detect IFN-γ KO Jurkat cells): Sample Type Dilution Ratio Content Wild Jurkat cells were stimulated with 50 ng/mL PMA and 1 μM ionomycin for 12 hours to detect cell culture supernatant undiluted 299pg/mL KO Jurkat cells were stimulated with 50 ng/mL PMA and 1 μM ionomycin for 12 hours to detect cell culture supernatant undiluted ND KO Jurkat cells were treated with 50 ng/mL PMA and 1 μM ionomycin for 12 hours and then cultured with 300 ng/mL Brefeldin A (BFA) for 3 hours. Add cell lysis buffer(Catalogue No.:E050), collect the lysate solution (total protein concentration measured by BCA assay: 2.23 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 IFN-γ KO Jurkat cells):

Sensitivität: 9.375 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 Interferon gamma

Target: Interferon gamma (IFNG)

Andere Bezeichnung: IFN-gamma

Hintergrund: Interferon gamma, IFN-gamma, Immune interferon, IFNG

UniProt: P01579

Pathways: Interferon-gamma Pathway, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Protein targeting to Nucleus, Autophagie