Analysis Note

Positive ControlMouse dorsal root nerve fibers

Application

Immunoblotting (1:1000)Frozen Sections (1:2000, fluorescence)

General description

This Anti-PGP9.5 (176-191) Rabbit pAb is validated for use in Immunoblotting, Frozen Sections for the detection of PGP9.5 (176-191).

Recognizes the ~26 kDa PGP9.5 protein in mouse dorsal root nerve fibers.

Rabbit polyclonal antibody supplied as undiluted serum. Recognizes the ~26 kDa PGP9.5 protein.

Immunogen

a synthetic peptide (ASSEDTLLKDAAKVCR) corresponding to amino acids 176-191 of PGP9

Legal Information

Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

PGP9.5 is also overexpressed in some cancers.r>r>Immunofluorescence protocolr>Care should be taken so that the incubation solutions do not evaporate. It is recommended that incubations be performed in a humidified chamber to prevent evaporation. Washing steps should be done in large volumes of phosphate buffered saline (PBS). r>1. Warm slides to room temperature. r>2. Re-hydrate with PBS for 10-15 min. r>3. Remove PBS and incubate in blocking buffer (PBS containing 3% normal donkey serum, 1% bovine serum albumin, 0.3% Triton^™ X-100 detergent, and 0.01% NaN₃, pH 7.45) for 1 h at room temperature. r>4. Dilute primary antibody in blocking buffer. Store diluted antibody at 4°C until use. r>5. Remove blocking buffer and add diluted primary antibody. r>6. Incubate with primary antibody at 4°C for 18-48 h. r>7. Remove primary antibody and wash three times for 10 min with PBS. r>8. Dilute secondary antibody in blocking buffer to appropriate dilution (follow manufacture′s recommendation). Store at 4°C until use. r>9. Incubate slides in diluted secondary antibody solution for one hour at room temperature.r>10. Remove secondary antibody and wash three times for 10 min with PBS.r>11. Coverslip slides with appropriate mounting media.

Caballero, O.L., et al. 2002. Oncogene21, 3003.Tezel, E., et al. 2000. Clin. Cancer Res.6, 4764.

Packaging

50 µL in Plastic ampoule

Physical form

Undiluted serum.

Reconstitution

Following initial thaw, aliquot and freeze (-20°C).

Warning

Toxicity: Standard Handling (A)