Suitable for Immunohistochemistry on: Frozen sections: 0.5 μg/mL (1/400)Paraffin sections: 1 μg/mL (1/200), Pretreatment not necessary. Also suitable for ELISA. Suggested positive control: Human tonsil. The antigen is a helical determinant of the native type IV collagen. It is stable toformaldehyde fixation and paraffin embedding. The antigen can be detected in allbasement membranes with the exception of the basement membrane of the cornealepithelium. 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, 0.05 % (v/v) Kathon CG, 5 mg/mL BSA
Preservative
Thimerosal (Merthiolate)
Precaution of Use
This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Prior to reconstitution store at 2-8 °C. Following reconstitution store the antibody at -20 °C.
Collagen IV is a major constituent of the basement membranes along with laminins, proteoglycans and enactins. It is a multimeric protein composed of 3 alpha subunits. These subunits are encoded by 6 different genes, alpha 1 through alpha 6, each of which can form a triple helix structure with 2 other subunits to form type IV collagen. It can form insoluble fibers with high tensile strength. Collagen IV is useful in detecting the loss of parts of basement membranes in carcinomas.Synonyms: COL4A1