Data Sheet

Data Sheet
CD3 T-Cell (M)
Concentrated and Prediluted Monoclonal Antibody
Control Number: 901-110-032415
Catalog Number:
CM 110 AK, BK, CK
PM 110 AA, H
IP 110 G10
OAI 110 T60
0.1, 0.5, 1.0 ml, concentrated
6.0, 25 ml, prediluted
10 ml, prediluted
60 tests, prediluted
1:50 -1:100
Renoir Red
Intended Use:
For In Vitro Diagnostic Use
CD3 T-Cell (M) [PS1] is a mouse monoclonal antibody that is intended for laboratory
use in the qualitative identification of CD3 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:
Monoclonal antibody to human CD3, when used in conjunction with other antibodies,
is regarded as a reliable pan T-cell antibody used in the immunophenotyping of
lymphomas in paraffin sections. Most T-cell lymphomas show positivity for CD3.
Notable exceptions include some of the more aggressive large T-cell lymphomas and
anaplastic large cell (Ki-1, CD30) lymphomas, which may not express detectable
antigen. CD3 immunoreactivity has also been reported in a minority of Reed- Sternberg
cells of Hodgkin's disease and in some histiocytic tumors. CD3 expression of
hemapoietic cells of the lymphoid, myeloid, and erythroid lineages in the human fetal
and embryonic liver is rare. In a study of 50 archived T-cell lymphomas UCHL-1 (a
monoclonal antibody to CD45RO) showed reactivity with 94% of cases, but lacked
absolute specificity for T-cells, especially in high-grade lymphomas. CD3 showed
reactivity with 80% of neoplastic cells, but with a higher specificity (1). When used in
conjunction, UCHL-1 and monoclonal CD3 identified the majority of T-cell
lymphomas in paraffin sections (2).
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; others not tested.
Clone: PS1
Isotype: IgG2a
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig concentration
Epitope/Antigen: CD3
Cellular Localization: Predominantly cell membrane. Some cytoplasmic
Positive Tissue Control: Tonsil or T-cell lymphoma
Known Applications:
Immunohistochemstry (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: Borg or 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.
Protocol Recommendations (intelliPATH and manual use) Cont'd:
Protein Block (Optional): Incubate for 5-10 minutes at RT with Biocare's Background
Primary Antibody: Incubate for 30-45 minutes at RT.
Probe: Incubate for 10 minutes at RT with a secondary probe.
Polymer: Incubate for 10-20 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 with hematoxylin. Rinse with deionized water. Apply Tacha's Bluing
Solution for 1 minute. Rinse with deionized water.
Technical Note:
This antibody has been optimized for use with Biocare's MACH 4 Universal HRPPolymer Detection and intelliPATH Universal HRP Detection Kit. 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.
intelliPATH™ Automated Slide Stainer:
IP110 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):
OAI110 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: CD3
Protocol Template (Description): Ms HRP Template 1
Dewaxing (DS Option): DS2
Antigen Retrieval (AR Option): AR1, high pH; 101°C
Reagent Name, Time, Temp.: CD3, 30 min., 25°C
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
Page 1 of 2
CD3 T-Cell (M)
Concentrated and Prediluted Monoclonal Antibody
Control Number: 901-110-032415
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 into contact with sensitive
areas, wash with copious amounts of water. (4)
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. Cabecadas JM, Isaacson PG. Phenotyping of T-cell lymphomas in paraffin sections-which antibodies? Histopathology. 1991 Nov;19(5):419-24.
2. Steward M, et al. Production and characterization of a new monoclonal antibody
effective in recognizing the CD3 T-cell associated antigen in formalin-fixed embedded
issue. Histopathology. 1997 Jan;30(1):16-22.
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-Fourth Edition
CLSI document M29-A4 Wayne, PA 2014.
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