Mouse Endostatin ELISA Kit

Mouse Endostatin ELISA Kit Features

Product Name            Mouse Endostatin ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   2.5 ng/mL
Assay Range                2.5-600 ng/mL
Reactivity                     Mouse
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               Col18a1
Gene ID                         12822
Swissprot ID               P39061

Recovery

Linearity

Standard Curve: Mouse Endostatin ELISA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

Endostatin is a naturally occurring, 20-kDa C-terminal fragment derived from type XVIII collagen.It is a broad-spectrum angiogenesis inhibitor and may interfere with the pro-angiogenic action of growth factors such as basic fibroblast growth factor (bFGF/FGF-2) and vascular endothelial growth factor (VEGF).

Endostatin suppresses angiogenesis through many pathways affecting both cell viability and movement. It represses cell cycle control and anti-apoptosis genes in proliferating endothelial cells, resulting in cell death. Endostatin blocks pro-angiogenic gene expression controlled by c-Jun N terminal kinase (JNK) by interfering with TNFα activation of JNK. It reduces the growth of new cells by inhibiting cyclin D1. As a result, cells arrest during G1 phase and enter apoptosis. Alteration of FGF signal transduction by endostatin inhibits the migration of endothelial cells through disruption of cell-matrix adhesions, cell-cell adhesions, and cytoskeletal reorganization. By binding integrin α5β1 on endothelia cells it inhibits the signaling pathways of Ras and Raf kinases and decreases ERK-1 and p38 activity. Endostatin binding and clustering of integrins causes co-localization with caveolin-1 and activates non-receptor tyrosine kinases of the Src family involved in the regulation of cell proliferation, differentiation, and mobility.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status 

Mouse Endostatin ELISA Kit is for Research Use Only.

• Manual
• Assay procedure
• MSDS