IL-6 ELISA Kit

Catalog No. ABIN7671737

Human IL-6 ELISA Kit, Colorimetric assay for quantification of Human IL-6.

Quick Overview for IL-6 ELISA Kit (ABIN7671737)

Target: IL-6 (IL6) (Interleukin 6 (IL6))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 4.688 pg/mL - 300 pg/mL

Application: ELISA

Sample Type: Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate

Grade: Quick

Product Details IL-6 ELISA Kit

Minimum Detection Limit: 4.688 pg/mL

Purpose: Human IL-6 (Interleukin 6) QuickTest ELISA Kit

Analytical Method: Quantitative

Specificity: Specifically binds with IL-6 , 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 human serum/plasma (EDTA, Citrate , heparin) (n=10) undiluted 1.2-6pg/mL Antigen-positive serum of COVID-19 patients (n=5) 1/2-1/10 dilution 162-670pg/mL Antigen-positive plasma (EDTA) of COVID-19 patients (n=12) 1/2-1/10 dilution 225-1237pg/mL Healthy human saliva (n=9) undiluted ND-12pg/mL Human peripheral blood mononuclear cells (2 x 10^6 cells/mL) were cultured with 5 %FBS+ 1640 + double antibody +10 μg PHA/mL, and the cell culture supernatant was detected after 48 hours 1/100 dilution 7.5 ng/mL 2. KO sample validation (Detect IL-6 KO A549 cells): Sample Type Dilution Ratio Content Wild A549 cells were stimulated with 20 ng/mL(h.IL-11b) for 24 hours to detect cell culture supernatant 1/20 dilution 1.23 ng/mL KO A549 cells were stimulated with 20 ng/mL(h.IL-11b) for 24 hours to detect cell culture supernatants undiluted ND KO A549 cells were treated with 20 ng/mL(H.IL-11b) for 24 hours and then cultured with 5 μg/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: 2.51 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-6 KO A549 cells):

Sensitivity: 0.5 pg/mL

Components:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Cap/Det Ab (Ready to use
• Red)
• TMB Substrate
• Sample Dilution Buffer (blue)
• 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: 50 μL

Assay Time: 1.5 h

Plate: Pre-coated

Protocol: 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 five times without immersion. Step 2: 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 3: Add 50 µL stop solution. Read at 450nm immediately and calculate.

Assay Procedure:

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 five times without immersion.
3. 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.).
4. 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

Target Details for IL-6

Target: IL-6 (IL6) (Interleukin 6 (IL6))

Alternative Name: IL-6

Background: Interleukin-6, IL-6, B-cell stimulatory factor 2, BSF-2, CTL differentiation factor, CDF, Hybridoma growth factor, Interferon beta-2, IFN-beta-2, IL6, IFNB2

UniProt: P05231

Pathways: TLR Signaling, Hormone Transport, Negative Regulation of Hormone Secretion, Myometrial Relaxation and Contraction, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Regulation of Carbohydrate Metabolic Process, Autophagy, Cell RedoxHomeostasis, Cancer Immune Checkpoints, Inflammasome