Endothelium antibody (Biotin)

Catalog No. ABIN114380

Product Details

Target: Endothelium

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: Biotin

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

Purification: Protein G Chromatography

Immunogen: Rat Peritoneal Macrophages Immunocyte Donor: BALB/c Spleen Fusion Partner: NSO/U

Clone: OX-43

Isotype: IgG1

Application Details

Application Notes: Flow Cytometry.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-Rat cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 1. 0 µg of this Ab per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Rat Strain: WistarCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 1. 0 µg/10e6 cellsIsotypic Control: Biotin Mouse IgG1. Cell Source Percentage of cells stained above control: Peritoneal Macrophages: 91. 5%Thymus: 8. 9%Results - Strain Distribution: Antibody Concentration Used: 1. 0 µg/10e6 cellsStrains Tested: Wistar, Brown Norway, Buffalo, Fischr 344Positive: Wistar, Brown Norway, Buffalo, Fischr 344Negative: none

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Target Details

Target: Endothelium

Background: The endothelium is located at the interface between the blood and the vessel wall. The cells are in close contact and form a slick layer that prevents blood cell interaction with the vessel wall as blood moves through the vessel lumen. The endothelium consists of simple squamous epithelium that lines the lumen of all blood vessels. It plays a critical role in the mechanics of blood flow, the regulation of coagulation, leukocyte adhesion, and vascular smooth muscle cell growth, and also serves as a barrier to the transvascular diffusion of liquids and solutes. For years the endothelium was thought of as an inert single layer of cells that passively allow the passage of water and other small molecules across the vessel wall. However, this dynamic tissue performs many other active functions, such as the secretion and modification of vasoactive substances and the contraction and relaxation of vascular smooth muscle.Synonyms: endothelial cells, endothelial marker