Cadherin 5 ELISA Kit

Catalog No. ABIN2859329

This Colorimetric ELISA kit is designed for the quantitative measurement of Human Cadherin 5.

Quick Overview for Cadherin 5 ELISA Kit (ABIN2859329)

Target: Cadherin 5 (CDH5)

Binding Specificity: AA 48-599

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 312-20.000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Product Details Cadherin 5 ELISA Kit

Minimum Detection Limit: 312 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human VE-Cadherin/CD144

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: NSO, D48-Q599

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: D48-Q599

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Tissue Specificity: Endothelial tissues and brain.

Plate: Pre-coated

Protocol: human VE-Cadherin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for VE-Cadherin has been precoated onto 96-well plates. Standards(NSO, D48-Q599) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for VE-Cadherin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human VE-Cadherin amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 20,000pg/mL, 10,000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL human VE-Cadherin standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human VE-Cadherin standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 2.21, Standard deviation: 0.095, CV(%): 4.3
• Sample 2: n=16, Mean(ng/ml): 8.45, Standard deviation: 0.313, CV(%): 3.7
• Sample 3: n=16, Mean(ng/ml): 14.4, Standard deviation: 0.691, CV(%): 4.8,
• Sample 1: n=24, Mean(ng/ml): 2.13, Standard deviation: 0.124, CV(%): 5.8
• Sample 2: n=24, Mean(ng/ml): 8.79, Standard deviation: 0.466, CV(%): 5.3
• Sample 3: n=24, Mean(ng/ml): 13.7, Standard deviation: 1.04, CV(%): 7.6

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for Cadherin 5

Target: Cadherin 5 (CDH5)

Alternative Name: CDH5

Background:

Protein Function: Cadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells, cadherins may thus contribute to the sorting of heterogeneous cell types. This cadherin may play a important role in endothelial cell biology through control of the cohesion and organization of the intercellular junctions. It associates with alpha-catenin forming a link to the cytoskeleton. Acts in concert with KRIT1 to establish and maintain correct endothelial cell polarity and vascular lumen. These effects are mediated by recruitment and activation of the Par polarity complex and RAP1B. Required for activation of PRKCZ and for the localization of phosphorylated PRKCZ, PARD3, TIAM1 and RAP1B to the cell junction. .

Background: Cadherin 5, type 2 or VE-cadherin also known as CD144, is encoded by the gene CDH5. It is a classical cadherin from the cadherin superfamily. This gene is mapped to mouse chromosome 8. The encoded protein is a calcium-dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. Functioning as a classic cadherin by imparting to cells the ability to adhere in a homophilic manner, the protein may play an important role in endothelial cell biology through control of the cohesion and organization of the intercellular junctions. It has found that PECAM1, VE-cadherin and VEGFR2 together are sufficient to confer responsiveness to flow in heterologous cells.

Synonyms: Cadherin-5,7B4 antigen,Vascular endothelial cadherin,VE-cadherin,CD144,CDH5,

Full Gene Name: Cadherin-5

Cellular Localisation: Cell junction . Cell membrane, Single-pass type I membrane protein . Found at cell-cell boundaries and probably at cell-matrix boundaries. KRIT1 and CDH5 reciprocally regulate their localization to endothelial cell-cell junctions.

Gene ID: 1003

UniProt: P33151

Pathways: Cell-Cell Junction Organization, Signaling Events mediated by VEGFR1 and VEGFR2