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Alpha-Methylacyl-CoA Racemase/P504S

IHC of AMACR on an FFPE Prostatic Adenocarcinoma Tissue

Description

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.

Antibody Type
Rabbit Monoclonal
Clone
RBT-AMACR
Isotype
IgG
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Prostatic Adenocarcinoma
Storage
Store at 2°-8°C
Stability
2 years

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).

Presentation

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.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5057
Prediluted
Ready-To-Use
3.0 ml
BSB 5058
Prediluted
Ready-To-Use
7.0 ml
BSB 5059
Prediluted
Ready-To-Use
15.0 ml
BSB 5060
Concentrated
1:50-1:200
0.1 ml
BSB 5061
Concentrated
1:50-1:200
0.5 ml
BSB 5062
Concentrated
1:50-1:200
1.0 ml
BSB 5063
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Jiang Z, et al. P504S Am J Surg Pathol. 2001;25:1397-1404
  2. Rubin MA, et al. JAMA. 2002;287:1662-1670
  3. Luo J, et al. Res. 2002; 62:2220-2226
  4. Beach R, et al. Am J Surg Pathol. 2002;26:1588-1596

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. 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).
  5. 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.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain