IL-1 beta ELISA Kit

Catalog No. ABIN7671658

The Human IL-1 beta ELISA Kit (ABIN7671658) is a Colorimetric ELISA Kit designed to quantify Human IL-1 beta.

Quick Overview for IL-1 beta ELISA Kit (ABIN7671658)

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 3.906 pg/mL - 250 pg/mL

Application: ELISA

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

Grade: Quick

Product Details IL-1 beta ELISA Kit

Minimum Detection Limit: 3.906 pg/mL

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

Analytical Method: Quantitative

Specificity: 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):

Sensitivity: 2.344 pg/mL

Components:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Cap/Det Ab (Ready to use
• blue)
• 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: 50 μL

Assay Time: 2 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 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.

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 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.

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-1 beta

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

Alternative Name: IL-1beta

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

UniProt: P01584

Pathways: NF-kappaB Signaling, Interferon-gamma Pathway, TLR Signaling, 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, Autophagy, Cancer Immune Checkpoints, Inflammasome