IL-3 ELISA Kit

Catalog No. ABIN1112646

This Human IL-3 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Human IL-3.

Quick Overview for IL-3 ELISA Kit (ABIN1112646)

Target: IL-3 (Interleukin 3 (IL-3))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6-1000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Tissue Lysate

Product Details IL-3 ELISA Kit

Minimum Detection Limit: 15.6 pg/mL

Purpose: For quantitative detection of IL-3 in human serum, body fluids, tissue lysate or cell culture supernate.

Analytical Method: Quantitative

Sensitivity: < 1 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human IL-3 antibody 2. Lyophilized Human IL-3 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human IL-3 antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-IL-3 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-IL-3 polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the IL-3 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of IL-3 can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Body fluids, tissue lysate or cell culture supernate: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C.
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C . Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for IL-3

Target: IL-3 (Interleukin 3 (IL-3))

Alternative Name: IL-3

Background: Interleukin 3 (IL-3) is a hematopoietic colony-stimulating factor that is capable of supporting the proliferation of a broad range of hematopoietic cell types. It is chemically synthesized by means of an automated peptide synthesizer and is shown to have the biological activities attributed to native IL-3. It is secreted by basophils and activated T cells to support growth and differentiation of T cells from the bone marrow in an immune response. It can improve the body's natural response to disease as part of the immune system. It acts by binding to the interleukin-3 receptor. IL3 infusion expands an early cell population that subsequently requires the action of a later-acting factor such as GMCSF to complete its development.

Pathways: JAK-STAT Signaling, Regulation of Carbohydrate Metabolic Process, Autophagy