AMACR (P504S) clone 13H4


Catalogue number: P504S-0.2

Clone13H4
IsotypeIgG
Product Type Primary Antibodies
Units0.2 ml
HostRabbit
Application IHC(C
P)
Background
The monoclonal Rabbit antibody directed against AMACR (P504S) shows predominant binding to prostate carcinoma tissue and only very faint or no binding to healthy benign prostate or benign prostate hyperplasia. Formaldehyde fixed and paraffin embedded tissue was investigated by Jiang et al. Among a total of 207 tissue samples all 137 carcinoma samples were P504S-positive, independent of Gleason-Score (V->VIII) (Jiang et al., 2001). Thus it may be possible to distinguish tumour epithelial cells from normal epithelial prostate tissue with help of this antibody. The expression of P504S in prostate is inversely related to that of high molecular weight cytokeratins as detected by clone 34ßE12. The majority (82,5%) of atypical adenomatous hyperplasia of prostate (n=40) reacted negative with this antibody, (82,5%), focal staining was observed in 10% and diffuse positive staining in 7,5% of cases (Yang et al., 2002). AMACR will also be found in neoplasm of other organs, e.g. colorectal carcinoma, mamma carcinoma, and renal cell carcinoma (Zhou et al., 2002). Alpha-Methylacyl-CoA Racemase (AMACR) has originally been isolated and characterised from rat liver (Schmitz et al., 1994, later also from human liver (Schmitz et al., 1995). It is a peroxisomal and mitochondrial enzyme, specifically involved in beta-oxidation of branched fatty acids and fatty acid metabolites. Interestingly this enzyme by cDNA-subtraction and microarray screening has also been detected in prostate carcinoma (Jiang et al., 2001). cDNA of human AMACR consists 1621 bases encoding a 382 AA protein. Human AMACR (P504S) protein, Alpha-Methylacyl CoA Racemase.

Positive Control
Prostate carcinoma

Pre-Treatment
In formaldehyde fixed tissue unmasking of antigen by heat pre-treatment is required. We recommend the application of our unmasking fluid G on glycol basis (Art. No. DE007) or EDTA solution pH 9 (Art.-No. DE006) approx. 30 min in pressure cooker or micro wave oven. Citrate buffer is also suitable, but gives less intensity in staining and changes the cell morphology more than the above mentioned unmasking methods.

Immunogen
Human recombinant AMACR (P504S) protein, whole sequence

Working Concentration
(liquid conc.) 1:80 – 1:200

Concentration
40 µg/ml

Incubation Time
60 min at RT

Secondary Reagents
Any secondary system containing anti-rabbit IgG may be used.

Product
Ion exchange chromatography purified antibody in PBS, BSA, sodium azide (0.09%)*. Use antibody dilution buffer (e.g. Art. No. PU002) containing sufficient protein and preservative.

Applications
IHC(C, P)

Cross Reactivity
Human

Storage
2-8°C

References
1. Schmitz W., Albers C., Fingerhut R., and Conzelmann E. (1995) Purification and characterization of an alpha-methylacyl-CoA racemase from human liver. Eur J. Biochem. 231(3); 815-822. 2. Babcook J.S., Leslie K.B., Olsen O.A., et al. (1996) A novel strategy for generating monoclonal antibodies from single isolated lymphocytes producing antibodies of defined specificities. Proc. Natl. Acad. Sci. USA 93; 7843-7848. 3. Xu J., Stolk J.A., Zhang X., et al. (2000) Identificatin of differentially expressed genes in human prostate cancer using subtraction and microarray. Cancer Res. 60; 1677-1682. 4. Ferdinandusse S., Denis S., IJ1st, et al. (2000) Subcellular localization and physiological role of alpha-methylacyl-coA racemase. J. Lipid Res. 41; 1890-1896. 5. Jiang Z., Woda B.A., Rock K.L., Xu Y., Savas L., et al. (2001) P504S a new molecular marker for the detection of prostate carcinoma. Am. J. Surg. Pathol. 25(11): 1397-1404.

Caution
*These antibodies are intended for in vitro research use only. They must not be used for clinical diagnostics and not for in vivo experiments in humans or animals. ** The preservative sodium azide is known to be poisonous and potentially hazardous to health. It should be handled only by trained staff. Despite of the product's low azide concentration it must be handled with care. Dispose according to regional rules!