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                                                                PIN Cocktail (P504S + p63)                                                                              9001:2000
                                                                           Prediluted Cocktail Antibody
                                                                            Control Number: 902-201-112910

Catalog Number:        PPM 201 AA, H
                                                                                             Pretreatment Protocol:
Description:           6.0, 25 ml, prediluted                                                Heat Retrieval Method:
Dilution:              Ready-to-use                                                          Retrieve sections under pressure using Biocare's Decloaking Chamber, followed by a
                                                                                             wash in distilled water. Alternatively, steam tissue sections for 45-60 minutes. Allow
Diluent:               N/A
                                                                                             solution to cool for 10 minutes then wash in distilled water.
Intended Use:                                                                                Avidin-Biotin Block is required:
For Research Use Only                                                                        1. Apply Avidin solution for 10 minutes and wash with TBS wash buffer.
                                                                                             2. Apply Biotin solution for 10 minutes and wash with TBS wash buffer.
Summary and Explanation:
                                                                                             Protein Block:
P504S is an enzyme in the ß-oxidation of branched-chain fatty acids. Expression of
P504S protein is found in prostatic adenocarcinoma but not in benign prostatic tissue.       Optional: Incubate for 10-15 minutes at RT with Biocare's Background Sniper.
It has also been found to stain premalignant lesions of the prostate: high-grade prostatic   Primary Antibody: Incubate for 30 minutes at RT.
intraepithelial neoplasia (PIN) and atypical adenomatous hyperplasia. P504S can be           Link: Incubate for 10 minutes at RT with a link.
used as a positive marker for PIN. It will be useful to confirm the diagnosis of small       Label: Incubate for 10 minutes at RT with a label.
foci of prostate carcinoma in needle biopsies. P504S stains the majority prostate            Chromogen:
cancer, however, P504S has been shown to stain many other types of carcinoma such            Incubate for 5 minutes at RT when using Biocare's DAB -OR- Incubate for 10-20
as hepatoma, breast carcinoma, pancreatic islet tumor and desmoplastic small round           minutes at RT when using Biocare's Vulcan Fast Red.
cell tumor (Miller and Tacha)1.                                                              Counterstain:
                                                                                             Counterstain with Hematoxylin. Rinse with deionized water. Apply Tacha's Bluing
p63 encodes for different isotypes able to either transactivate p53 reporter genes           solution for 1 minute. Rinse with deionized water.
(TAp63) or act as p53-dominant-negatives. p63 is detected in prostate basal epithelial
                                                                                             Technical Note:
nuclei in normal prostate, however, is negative in malignant tumors of the prostate
gland. Thus p63 is useful as a differential marker for benign and malignant tumors of        This antibody has been standardized with Biocare's 4 plus detection system. It can also
prostate gland and can be useful as a negative marker.                                       be used on an automated staining system and with other Biocare polymer detection
                                                                                             kits. Use TBS buffer for washing steps.
The combination of P504S and p63 (PIN cocktail) may be extremely useful for                  Performance Characteristics:
diagnosing prostatic intraepithelial neoplasia (PIN), especially in difficult and limited    The optimum antibody dilution and protocols for a specific application can vary. These
tissue cases. This antibody cocktail may eliminate the need for HMW CK [34ßE12].             include, but are not limited to: fixation, heat-retrieval method, incubation times, tissue
P504S stains cytoplasm in prostate adenocarcinoma and atypical adenomatous                   section thickness and detection kit used. Due to the superior sensitivity of these unique
hyperplasia; p63 stains nuclei in normal (negative marker) and benign prostate glands.       reagents, the recommended incubation times and titers listed are not applicable to other
In this instance, the PIN Cocktail becomes a double stain procedure. (A goat anti-           detection systems, as results may vary. The data sheet recommendations and protocols
mouse and rabbit universal detection kit must be used).                                      are based on exclusive use of Biocare products. Ultimately, it is the responsibility of
                                                                                             the investigator to determine optimal conditions. These products are tools that can be
Source: Rabbit polyclonal and Mouse monoclonal                                               used for interpretation of morphological findings in conjunction with other diagnostic
                                                                                             tests and pertinent clinical data by a qualified pathologist.
Species Reactivity: Human; others not tested.
                                                                                             Quality Control:
Clone: BC4A4 (p63)
                                                                                             Refer to NCCLS Quality Assurance for Immunocytochemistry approved guidelines,
Isotype: Mouse IgG2a (p63)                                                                   December 1999 MM4-A Vol.19 No.26 for more information about tissue controls.
Total Protein Concentration: ~10 mg/ml                                                       Precautions:
Epitope/Antigen: P504S + p63                                                                 This antibody contains less than 0.1% sodium azide. Concentrations less than 0.1% are
                                                                                             not reportable hazardous materials according to U.S. 29 CFR 1910.1200, OSHA
Cellular Localization: Granular cytoplasmic (P504S) and nuclear (p63)                        Hazard communication and EC Directive 91/155/EC.
Positive Control: Prostatic intraepithelial neoplasia or prostate cancer                     Sodium azide (NaN3) used as a preservative is toxic if ingested. Sodium azide may
                                                                                             react with lead and copper plumbing to form highly explosive metal azides. Upon
Normal Tissue: N/A
                                                                                             disposal, flush with large volumes of water to prevent azide build-up in plumbing.
Abnormal Tissue: Prostate intraepithelial neoplasm                                           (Center for Disease Control, 1976, National Institute of Occupational Safety and
Known Applications:                                                                          Health, 1976)
Immunohistochemistry (formalin-fixed paraffin-embedded tissues)                              Specimens, before and after fixation, and all materials exposed to them should be
                                                                                             handled as if capable of transmitting infection and disposed of with proper precautions.
Supplied As: Buffer with protein carrier and preservative.                                   Never pipette reagents by mouth and avoid contacting the skin and mucous membranes
Storage and Stability:                                                                       with reagents and specimens. If reagents or specimens come in contact with sensitive
Store at 2ºC to 8ºC. Do not use after expiration date printed on vial. If reagents are       areas, wash with copious amounts of water.
stored under conditions other than those specified in the package insert, they must be       Microbial contamination of reagents may result in an increase in nonspecific staining.
verified by the user. Diluted reagents should be used promptly; any remaining reagent        Incubation times or temperatures other than those specified may give erroneous results.
should be stored at 2ºC to 8ºC.                                                              The user must validate any such change. The MSDS is available upon request.
Protocol Recommendations
Peroxide Block:
Block for 5 minutes with Biocare's Peroxidazed 1.
Pretreatment Solution (recommended): Reveal

                                                                                      Page 1 of 2
                                                              PIN Cocktail (P504S + p63)                 9001:2000
                                                                      Prediluted Cocktail Antibody
                                                                       Control Number: 902-201-112910

Follow the antibody specific protocol recommendations according to data sheet
provided. If atypical results occur, contact Biocare's Technical Support at
Limitations and Warranty:
There are no warranties, expressed or implied, which extend beyond this description.
Biocare is not liable for property damage, personal injury, or economic loss caused by
this product.
1. Tacha DE, Miller RT. Use of p63/P504S monoclonal antibody cocktail in
immunohistochemical staining of prostate tissue. Appl Immunohistochem Mol
Morphol. 2004 Mar;12(1):75-8.Biocare Medical, Walnut Creek, California.
2. Jiang Z, Woda BA et al. P504S: a new molecular marker for the detection of
prostate carcinoma. Am J Surg Pathol 2001 Nov;25(11):1397-404
3. Signoretti S, Waltregny D et al. p63 is a prostate basal cell marker and is required
for prostate development. Am J Pathol 2000 Dec; 157(6):1769-75.
4. Beach R, Gown AM et al. P504S immunohistochemical detection in 405 prostatic
specimens including 376 18-gauge needle biopsies. Am J Surg Pathol 2002 Dec;26
5. Luo J, Zha S, Gage WR, Dunn TA, Hicks JL, Bennett CJ, Ewing CM, Platz EA,
Ferdinandusse S, Wanders RJ, Trent JM, Isaacs WB, De Marzo AM. Alpha-
methylacyl-CoA racemase: a new molecular marker for prostate cancer. Cancer Res
2002 Apr 15;62(8):2220-6
6. Center for Disease Control Manual. Guide: Safety Management, NO. CDC-22,
Atlanta, GA. April 30, 1976 "Decontamination of Laboratory Sink Drains to Remove
Azide Salts."
7. National Committee for Clinical Laboratory Standards (NCCLS). Protection of
laboratory workers from infectious diseases transmitted by blood and tissue; proposed
guideline. Villanova, PA 1991;7(9). Order code M29-P.

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