Detects human granulocytes, monocytes and platelets. X-6 detects an antigen of 154-180 kDa by immunoprecipitation. Antigen distribution on Isolated cells and Tissue sections: Granulocytes, monocytes. Positive on blood monocyte-derived macrophages days 0-28. Dull staining with 95 % cord blood CD34 cells. Weak reaction on resting neutrophils. Staining increase with maturation of granulocytes. Treatment with fMLP, TNF, GM-CSF and temperature causes postive shift with granulocytes and monocytes. PMA treatment of granulocytes leads to decrease in no. of positive cells. X-6 is positive on lining cells in different infiltrations and endothelial cells in the synovial membrane of RA patients. The staining pattern is similar to CD34 in these patients.
ITGA5
Reactivity: Human
WB, IF
Host: Rabbit
Polyclonal
unconjugated
Application Notes
Suitable for Immunohistochemistry on frozen sections (2 μg/mL (1: 100) and FACS (use 10 μLstock solution to label 10^6 cells). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain with Mayer's hemalum.
Restrictions
For Research Use only
Reconstitution
Restore with 0.5 mL distilled water.
Concentration
0.2 mg/mL
Buffer
PBS, pH 7.2 containing 10 mg/mL bovine serum albumin (BSA) as a stabilizer and 0.01 % Thimerosal as a preservative.
Preservative
Thimerosal (Merthiolate)
Precaution of Use
This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
The X-6 antibody is useful for the detection of the fibronectin receptor, or CD49e. CD49e is a component of an adhesion receptor heterodimer (α5β1) to fibronectin and invasin. α5β1 recognizes an Arg-Gly-Asp sequence in fibronectin. Binding of fibronectin through α5β1 is important for cell survival and induces apoptosis, particularly in hemopoietic cells.Synonyms: CD49 antigen-like family member E, FNRA, Fibronectin receptor subunit alpha, Integrin alpha-5, Integrin alpha-F, VLA-5, VLA5