
AMACR (P504S) is an acronym for the protein alpha-methylacyl CoA racemase that helps to metabolize certain fatty acids within the body. AMACR has been recently described as a prostate cancer-specific gene that encodes a protein involved in the beta-oxidation of branched chain fatty acids. Expression of AMACR protein is found in Prostatic Adenocarcinoma but not in benign prostatic tissue. It stains premalignant lesions of the prostate: High-Grade Prostatic Intraepithelial Neoplasia (PIN) and Atypical Adenomatous Hyperplasia. Several studies have suggested that AMACR can be used as a prostate cancer biomarker.
High expression of AMACR (P504S) protein is usually found in Prostatic Adenocarcinoma but not in benign prostatic tissue by immunohistochemical staining in paraffin-embedded tissues. Using AMACR as a positive marker along with basal-cell staining (34 beta E12 or p63) as a negative marker could help to confirm the diagnosis of small foci of Prostate Carcinoma on needle biopsies.
For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).
P504S/AMACR is a rabbit monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
- Jiang Z, et al. P504S Am J Surg Pathol. 2001;25:1397-1404
- Rubin MA, et al. JAMA. 2002;287:1662-1670
- Luo J, et al. Res. 2002; 62:2220-2226
- Beach R, et al. Am J Surg Pathol. 2002;26:1588-1596
Recommended Immunohistochemical Protocol
- Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
- Air dry for 2 hours at 58° C.
- Deparaffinize, dehydrate and rehydrate tissues.
- Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
- Any of three heating methods may be used:
- Electric Pressure Cooker
Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature. - Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes. - Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
- Electric Pressure Cooker
- After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
- Wash slides with IHC wash buffer or DI water.
- Continue IHC staining protocol.
Immunohistochemical Protocol
ImmunoDetector
(AP or HRP)
(AP or HRP)