ICAM1 ELISA Kit

Catalog No. ABIN1112623

Rat ICAM1 ELISA Kit Colorimetric assay for quantification of Rat ICAM1.

Quick Overview for ICAM1 ELISA Kit (ABIN1112623)

Target: ICAM1 (Intercellular Adhesion Molecule 1 (ICAM1))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

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

Product Details ICAM1 ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: For quantitative detection of ICAM-1 in rat serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 2 pg/mL

Components: 1. One 96-well plate pre-coated with anti-rat ICAM-1 antibody 2. Lyophilized rat ICAM-1 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-rat ICAM-1 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-ICAM-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-ICAM-1 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 ICAM-1 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of ICAM-1 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 lysates and cell culture supernatants: 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 1500 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with citrate, heparin or EDTA as the anticoagulant. Centrifuge for 15min at 1500 x g within 30 min of collection. Analyze immediately or aliquot and store frozen 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 ICAM1

Target: ICAM1 (Intercellular Adhesion Molecule 1 (ICAM1))

Alternative Name: ICAM-1 / CD54

Target Type: Viral Protein

Background: ICAM-1 (Intercellular Adhesion Molecule 1) also known as CD54 (Cluster of Differentiation 54) is a cell surface glycoprotein which is typically expressed on endothelial cells and cells of the immune system. It binds to integrins of type CD11a / CD18, or CD11b / CD18 and is also exploited by rhinovirus as a receptor. ICAM-1 is a member of the immunoglobulin superfamily, it can be induced by interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFĮ) and is expressed by the vascular endothelium, macrophages, and lymphocytes.It is also an endothelial- and leukocyte-associated transmembrane protein long known for its importance in stabilizing cell-cell interactions and facilitating leukocyte endothelial transmigration. More recently, ICAM-1 has been characterized as a site for the cellular entry of human rhinovirus.

Pathways: Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization, Carbohydrate Homeostasis, Regulation of Leukocyte Mediated Immunity, Thromboxane A2 Receptor Signaling