Synaptophysin

Synaptophysin
Synaptophysin
Prediluted Mouse Monoclonal Antibody
Control Number: 901-371IP-042313
Catalog Number:
IP 371 G10
Description:
10 ml, predilute
Intended Use:
For In Vitro Diagnostic Use
Synaptophysin [27G12] is a mouse monoclonal antibody that is intended for
laboratory use in the qualitative identification of synaptophysin protein by
immunohistochemistry (IHC) in formalin-fixed paraffin-embedded (FFPE) human
tissues. The clinical interpretation of any staining or its absence should be
complemented by morphological studies using proper controls and should be
evaluated within the context of the patient's clinical history and other diagnostic tests
by a qualified pathologist.
Summary and Explanation:
This monoclonal antibody [clone 27G12] is a reliable marker for neuroendocrine and
neuronal neoplasms, as well as their normal counterparts (1-2). Studies have shown
Synaptophysin has a higher degree of specificity than NSE, but is often used in
conjunction with CD56, NSE and chromogranin. This clone aids in providing sharper
staining when compared to other rabbit synaptophysin antibodies.
Pheochromocytoma stained with mouse Synaptophysin antibody
Principle of Procedure:
Antigen detection in tissues and cells is a multi-step immunohistochemical process.
The initial step binds the primary antibody to its specific epitope. After labeling the
antigen with a primary antibody, a secondary antibody is added to bind to the primary
antibody. An enzyme label is then added to bind to the secondary antibody; this
detection of the bound antibody is evidenced by a colorimetric reaction.
Source: Mouse monoclonal
Protocol Recommendations:
Pretreatment Solution (recommended): Reveal or Borg
Pretreatment Protocol:
Heat Retrieval Method:
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
solution to cool for 10 minutes then wash in distilled water.
Peroxide Block: Block for 5 minutes at RT.
Protein Block (Optional): Incubate for 5-10 minutes at RT.
Primary Antibody: Incubate for 45 minutes at RT.
Secondary: Incubate for 10 minutes at RT.
Tertiary: Incubate for 20 minutes at RT.
Chromogen: Incubate for 5 minutes with DAB at RT.
Counterstain:
1. Rinse with deionized water.
2. Incubate for 5 minutes with automated Hematoxylin.
3. Rinse with TBS Buffer for 1 minute followed by a rinse with deionized water.
Staining Procedure:
Biocare protocols have been standardized using in-house antibodies, detection and
accessory reagents for use on the intelliPATH automated stainer. Recommended
staining protocols are specified in the datasheet of the antibody of interest. Preoptimized intelliPATH protocols with preset parameters can be displayed, printed and
edited according to the procedure in the Operator's Manual. Refer to the Operator's
Manual for additional instruction to navigate intelliPATH software and stainer. Use
TBS for washing steps unless otherwise specified.
Limitations:
The optimum antibody dilution and protocols for a specific application can vary. These
include, but are not limited to: fixation, heat-retrieval method, incubation times, tissue
section thickness and detection kit used. Due to the superior sensitivity of these unique
reagents, the recommended incubation times and titers listed are not applicable to other
detection systems, as results may vary. The data sheet recommendations and protocols
are based on exclusive use of Biocare products. Ultimately, it is the responsibility of
the investigator to determine optimal conditions. The clinical interpretation of any
positive or negative staining should be evaluated within the context of clinical
presentation, morphology and other histopathological criteria by a qualified
pathologist. The clinical interpretation of any positive or negative staining should be
complemented by morphological studies using proper positive and negative internal
and external controls as well as other diagnostic tests
Quality Control:
Refer to CLSI Quality Standards for Design and Implementation of
Immunohistochemistry Assays; Approved Guideline-Second edition (I/LA28-A2)
CLSI Wayne, PA USA (www.clsi.org). 2011
Species Reactivity: Humans, others not tested
Clone: 27G12
Isotype: IgG1
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig concentration.
Epitope/Antigen: Synaptophysin
Cellular Localization: Cytoplasmic
Positive Control: Pancreas, colon, small cell lung carcinoma
Known Applications:
Immunohistochemistry (formalin-fixed paraffin-embedded tissues)
Supplied As: Buffer with protein carrier and preservative
Storage and Stability:
Store at 2ºC to 8ºC. Do not use after expiration date printed on vial. If reagents are
stored under conditions other than those specified in the package insert, they must be
verified by the user.
Precautions:
1. 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
Hazard communication and EC Directive 91/155/EC. 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 disposal, flush with large
volumes of water to prevent azide build-up in plumbing. (Center for Disease Control,
1976, National Institute of Occupational Safety and Health, 1976) (3)
2. 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.
Never pipette reagents by mouth and avoid contacting the skin and mucous membranes
with reagents and specimens. If reagents or specimens come in contact with sensitive
areas, wash with copious amounts of water. (4)
3. Microbial contamination of reagents may result in an increase in nonspecific
staining.
4. Incubation times or temperatures other than those specified may give erroneous
results. The user must validate any such change.
5. Do not use reagent after the expiration date printed on the vial.
6. The MSDS is available upon request and is located at http://biocare.
net/support/msds/.
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Synaptophysin
Prediluted Mouse Monoclonal Antibody
Control Number: 901-371IP-042313
Troubleshooting:
Follow the antibody specific protocol recommendations according to data sheet
provided.
If atypical results occur, contact Biocare's Technical Support at
1-800-542-2002.
References:
1. Chejfec G, Falkmer S, Grimelius L, Jacobsson B, Rodensjo M, Wiedenmann B,
Franke WW, Lee I, Gould VE. Synaptophysin. A new marker for pancreatic
neuroendocrine tumors. Am J Surg Pathol 1987 Apr;11(4):241-247.
2. Wiedenmann B, Franke WW, Kuhn C, Moll R, Gould VE. Synaptophysin: a marker
protein for neuroendocrine cells and neoplasms. Proc Natl Acad Sci U S A 1986
May;83(10):3500-3504.
3. 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."
4. Clinical and Laboratory Standards Institute (CLSI). Protection of Laboratory
workers from occupationally Acquired Infections; Approved guideline-Third Edition
CLSI document M29-A3 Wayne, PA 2005.
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