Data Sheet

Data Sheet
Concentrated and Prediluted Monoclonal Antibody
Control Number: 901-048-052215
Catalog Number:
CM 048 A, C
PM 048 AA
IP 048 G10
OAI 048 T60
0.1, 1.0 ml, concentrated
6.0 ml, prediluted
10 ml, prediluted
60 tests, prediluted
Da Vinci Green
Intended Use:
For In Vitro Diagnostic Use
Vimentin [V9] is a mouse monoclonal antibody that is intended for laboratory use in
the qualitative identification of vimentin 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:
Vimentin recognizes a 57-60 kDa protein, which is identified as vimentin. It shows no
cross-reactivity with other closely related intermediate filament proteins such as desmin
and GFAP. Studies have shown that Vimentin is the main intermediate filament protein
in mesenchymal cells, and therefore is of value in the differential diagnosis of
undifferentiated neoplasms including melanoma and sarcoma. Vimentin can also serve
as an internal control for formalin-fixed tissues that are over-fixed.
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. A secondary antibody
may be applied to bind the primary antibody, followed by an enzyme labeled polymer;
or an enzyme labeled polymer may be applied directly to bind the primary antibody.
The detection of the bound primary antibody is evidenced by an enzyme-mediated
colorimetric reaction.
Source: Mouse monoclonal
Species Reactivity: Human
Clone: V9
Isotype: IgG1/kappa
Total Protein Concentration:~10 mg/ml. Call for lot specific Ig concentration.
Epitope/Antigen: Vimentin
Cellular Localization: Variable
Positive Control: Melanoma
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. Diluted reagents should be used promptly; any remaining reagent
should be stored at 2ºC to 8ºC.
Protocol Recommendations (intelliPATH and manual use):
Peroxide Block: Block for 5 minutes with Biocare's Peroxidazed 1.
Pretreatment Solution (recommended): Reveal
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.
Protein Block (Optional): Incubate for 5-10 minutes at RT with Biocare's Background
Primary Antibody: Incubate for 15-30 minutes at RT.
Probe: Incubate for 7-10 minutes at RT with a secondary probe.
Protocol Recommendations (intelliPATH and manual use) Cont'd:
Polymer: Incubate for 7-10 minutes at RT with a tertiary polymer.
Chromogen: Incubate for 5 minutes at RT with Biocare's DAB - OR - Incubate for 5-7
minutes at RT with Biocare's Warp Red.
Counterstain: Counterstain with hematoxylin. Rinse with deionized water. Apply
Tacha's Bluing Solution for 1 minute. Rinse with deionized water.
intelliPATH™ Automated Slide Stainer:
IP048 is intended for use on the intelliPATH™ Automated Slide Stainer. Refer to the
intelliPATH Automated Slide Stainer manual for specific instructions on its use. When
using the intelliPATH, peroxide block with intelliPATH Peroxidase Blocking Reagent
(IPB5000) may be performed following heat retrieval.
Protocol Recommendations (ONCORE Automated Slide Staining System):
OAI048 is intended for use with the ONCORE Automated Slide Staining System.
Refer to the ONCORE Automated Slide Staining System User Manual for specific
instructions on its use. Protocol parameters in the ONCORE Automated Slide Stainer
Protocol Editor should be programmed as follows:
Protocol Name: Vimentin
Protocol Template (Description): Ms HRP Template 1
Dewaxing (DS Option): DS2
Antigen Retrieval (AR Option): AR2, low pH; 90°C
Reagent Name, Time, Temp.: Vimentin, 30 min., 25°C
Technical Note:
This antibody has been optimized for use with Biocare's MACH 4 Universal HRPPolymer Detection, intelliPATH Universal HRP Detection Kit and ONCORE HRP
Detection. Other Biocare polymer detection kits may be used; however, users must
validate incubation times and protocols for their specific application. Use TBS for
washing steps.
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 ( 2011.
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
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Concentrated and Prediluted Monoclonal Antibody
Control Number: 901-048-052215
Precautions Cont'd:
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) (4)
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. (5)
3. Microbial contamination of reagents may result in an increase in nonspecific
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 SDS is available upon request and is located at
Follow the antibody specific protocol recommendations according to data sheet
If atypical results occur, contact Biocare's Technical Support at
1. Urban K, Hewicker-Trautwein M. Fixation-dependent vimentin immunoreactivity of
mono- and polyclonal antibodies in brain tissue of cattle, rabbits, rats and mice. Acta
Histochem. 1994 Dec;96(4):365-77.
2. Battifora H. Assessment of antigen damage in immunohistochemistry. The vimentin
internal control. Am J Clin Pathol. 1991 Nov;96(5):669-71.
3. Carbone A, et al. Anti-vimentin antibody reactivity with Reed-Sternberg cells of
Hodgkin's disease. Virchows Arch A Pathol Anat Histopathol. 1990;417(1):43-8.
4. 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."
5. Clinical and Laboratory Standards Institute (CLSI). Protection of Laboratory
Workers from Occupationally Acquired Infections; Approved Guideline-Fourth Edition
CLSI document M29-A4 Wayne, PA 2014.
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