Blood Vessel Staining Kit; 1 k

Blood Vessel Staining Kit

Chemicon (Millipore)

Reagents and materials supplied in the Blood Vessel Staining Kit are sufficient for staining 100 tissue sections and when stored at 2-8ºC is stable up to the expiration date. ECM590 detects vWF and CD31 in human paraffin embedded tissues. In rat or mouse tissues only the vWF antibody is effective in paraffin; the CD31 antibody does not react with rodent CD31.

Background:

Angiogenesis and tumor-associated neovascularization are complex processes by which new blood vessels are formed from pre-existing vasculature. Several studies implicate tumor-associated blood vessel formation as a central part in the process of growth, invasion and metastasis of malignancies (Rak et al., 1995; Jahroudi & Greenberger, 1995; Saphir, 1997).

The most widely used endothelial cell markers for studying angiogenesis/neovascularization are von Willebrand Factor (Factor VIII Related Antigen) and CD31 (PECAM-1). von Willebrand Factor is synthesized by endothelial cells, causing adhesion of platelets to injured vessel walls and functions as a carrier and stabilizer for coagulation of Factor VIII. CHEMICON's von Willebrand Factor antibody reacts specifically with the endothelial cells of blood vessels and is a useful marker for the identification of endothelial lineage in tumors. It is important to note that not all endothelial cells synthesize von Willebrand factor, therefore a small percentage of these vascular-rich tumors might not stain with this antibody. CD31 or platelet endothelial adhesion molecule (PECAM-1), is a type I integral membrane glycoprotein and a member of the immunoglobulin super family of cell surface receptors. It is strongly expressed by all endothelial cells and weakly by several types of leucocytes (Parums et al., 1990). According to several studies, staining of blood vessels with CD31 antibodies has shown to be suitable for the identification of angiogenesis in several types of malignancies such as lung (Giatromanolaki et al., 1996), breast (Horak et al., 1992) and colorectal cancer (Takebayashi et al., 1996). CHEMICON's monoclonal antibody to CD31 is very specific for the staining of vascular endothelial cells in normal and cancerous tissue.

CHEMICON's Blood Vessel Staining Kit is a very sensitive immunohistochemical tool for the detection of vascular endothelial cells, containing both anti-von Willebrand Factor and anti-CD31 primary antibodies and is applicable to a variety of species. This immunoperoxidase kit provides a simple, all-in-one method for the staining of frozen and paraffin-embedded sections in human (both CD31 and VWF) or rat and mouse (VWF only) tissues

• Immunohistochemistry
• Immunohistochemistry (Paraffin)

ECM590 detects vWF and CD31 in human paraffin embedded tissues. In rat or mouse tissues only the vWF antibody is effective in paraffin; the CD31 antibody does not react with rodent CD31.

Human

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

• Apoptosis Assays
• Cancer Kits
• Cell Based Assays

• Rabbit anti-vWF Polyclonal Antibody (Part No. 21540): One vial containing 100 μg at 1 mg/mL.
• Mouse anti-CD31 Monoclonal Antibody (Part No. 90214): One vial containing 100 μL at 15 mg/mL (total protein including IgG and Carrier protein).
• Goat anti-Rabbit Secondary Antibody (Part No. 21537): One 15 mL ready-to-use solution containing biotinylated goat anti-rabbit IgG in PBS with a carrier protein.
• Goat anti-Mouse Secondary Antibody (Part No. 21538): One 15 mL ready-to-use solution containing biotinylated goat anti-mouse IgG in PBS with a carrier protein.
• Blocking Reagent (Part No. 20783): One 15 mL ready-to-use solution containing normal goat serum in PBS with a carrier protein.
• Streptavidin-HRP (Part No. 90215): One 15 ml ready-to-use solution of Streptavidin-HRP diluted in Tris-buffered saline (TBS).
• DAB Chromogen A (Part No. 90216): One 5 mL bottle of 3, 3' Diaminobenzidine diluted in TBS.
• DAB Chromogen B (Part No: 90217): Two 12 mL bottles of hydrogen peroxide diluted in distilled water.
• 20X Rinse Buffer (Part No. 90218): One 275 mL bottle of 20X TBS.

Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date. Do not store kit components or perform staining in strong light, such as direct sunlight.

Chromogenic

sufficient for 100 sections

1. Tissue Sections/Slides: Paraffin-embedded or frozen sections can be used with this kit.

2. 100% Ethanol.

3. Xylene.

4. Hematoxylin: If counterstaining if required.

5. Solutions used for epitope/antigen retrieval: For example, 10 mM Sodium Citrate (pH 6.0) solution.

6. Hydrogen Peroxide: Needed if quenching of endogenous peroxidase activity is required.

7. Mounting Fluid: Aqueous or Xylene-based.

8. Coverslips.

9. Microscope.