Interferon gamma ELISA Kit

Catalog No. ABIN5520846

Human Interferon gamma ELISA Kit, Colorimetric assay for quantification of Human Interferon gamma, that has been mentioned in 1 publication.

Quick Overview for Interferon gamma ELISA Kit (ABIN5520846)

Target: Interferon gamma (IFNG)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.625 pg/mL - 1000 pg/mL

Application: ELISA

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

Product Details Interferon gamma ELISA Kit

Minimum Detection Limit: 15.625 pg/mL

Purpose: Human IFN-γ (Interferon gamma) ELISA Kit

Analytical Method: Quantitative

Specificity: 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. (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=21) 1/2 dilution ND-22pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=21) 1/2 dilution ND-16pg/mL Human peripheral blood lymphocyte were cultured with 5 %FBS + 1640 + double antibody, and the cell culture supernatant was detected after 12 hours. 1/2 dilution ND 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. 1/2 dilution 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. 1/5 dilution 1.2 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 1/2 dilution 296pg/mL KO Jurkat cells were stimulated with 50 ng/mL PMA and 1 μM ionomycin for 12 hours to detect cell culture supernatant 1/2 dilution 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. 1/2 dilution 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):

Sensitivity: 9.375 pg/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

References for Interferon gamma ELISA Kit (ABIN5520846)

Kim, Kim, Kim: "The effects of a single bout pilates exercise on mRNA expression of bone metabolic cytokines in osteopenia women." in: Journal of exercise nutrition & biochemistry, Vol. 18, Issue 1, pp. 69-78, (2015) (PubMed).

Target Details for Interferon gamma

Target: Interferon gamma (IFNG)

Alternative Name: IFNgamma

Background: 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, Autophagy