IL-6 ELISA Kit

Catalog No. ABIN8090744

This Colorimetric ELISA kit is designed for the quantitative measurement of Mouse IL-6.

Quick Overview for IL-6 ELISA Kit (ABIN8090744)

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

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 3.125 pg/mL - 200 pg/mL

Application: ELISA

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

Grade: High Sensitivity

Product Details IL-6 ELISA Kit

Minimum Detection Limit: 3.125 pg/mL

Purpose: Mouse IL-6 (Interleukin 6) 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. (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 Normal mouse serum/plasma (EDTA, Citrate , heparin) (n=16) 1/2 dilution ND-42pg/mL Normal mouse were intraperitoneally injected with 50 μg LPS, and the serum was detected 12 hours later (n=3) 1/10-1/50 dilution 819-1906pg/mL Mouse spleen tissue homogenates 1/2 dilution 133pg/1 mg (total protein) RAW 264.7 cells were cultured with 5 %FBS + 1640 + double antibody for 7 hours 1/2 dilution 26pg/mL RAW 264.7 cells were cultured with 5 %FBS+ 1640 + double antibody +100 ng/mL LPS, and the cell culture supernatant was detected after 7 hours 1/10 dilution 687pg/mL 2. KD sample validation (Detect IL-6-KD RAW 264.7 cells): Sample Type Dilution Ratio Content Wild RAW 264.7 cells were stimulated with 100 ng/mL LPS for 7 hours to detect cell culture supernatant 1/10 dilution 870pg/mL RAW 264.7 cells (+siRNA) were stimulated with 100 ng/mL LPS for 7 hours to detect cell culture supernatants 1/2 dilution ND RAW 264.7 cells (+siRNA) were treated with 100 ng/mL LPS for 7 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: 1.69 mg/mL) to detect. 1/2 dilution 21pg/mg(total protein) Note:ND is lower than the sensitivity of the kit and was not detected 3. Antibody by WB KD validation (Detect IL-6-KO RAW 264.7 cells):

Sensitivity: 1.875 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

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: P08505

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