Leukocyte Common Antigen (LCA) Cocktail
Leukocyte Common Antigen (LCA) Cocktail
Concentrated and Prediluted Cocktail Antibody
Control Number: 901-016-010716
Catalog Number:
CM 016 AK, BK, CK
PM 016 AA
IP 016 G10
OAI 016 T60
Description:
0.1, 0.5, 1.0 ml, concentrated
6.0 ml, prediluted
10 ml, prediluted
60 tests, prediluted
Dilution:
1:100-1:200
Ready-to-use
Ready-to-use
Ready-to-use
Diluent:
Van Gogh Yellow
N/A
N/A
N/A
Intended Use:
For In Vitro Diagnostic Use
Leukocyte Common Antigen (LCA) Cocktail [PD7/26+2B11] is a mouse monoclonal
antibody cocktail that is intended for laboratory use in the qualitative identification of
leukocyte cellular surface marker protein CD45 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:
Studies have shown CD45 recognizes an antigen found on lymphoid cells. Most
neoplastic B-cells and T-cells stain positively in leukemia and in non-Hodgkin's
lymphomas; whereas most neoplastic myeloid and erythroid cells are negative (1-3).
Studies have also shown it is unreactive with epithelium and connective tissues. The
PD7/26 and 2B11 antibody was included in the 4th International Workshop and was
designated as CD45. It belongs to an LCA family of glycoproteins with molecular
weights of 180, 190, 205 and 220. It is well suited for formalin-fixed paraffinembedded tissues and is used as a pan lymphocyte screener for lymphoma (1-3).
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: PD7/26 and 2B11
Isotype: IgG1/kappa
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig concentration.
Epitope/Antigen: CD45 (Leukocyte Common Antigen)
Cellular Localization: Cell surface
Positive Control: Tonsil or lymphoma
Known Applications:
Immunohistochemistry (formalin-fixed paraffin-embedded tissues)
Supplied As: Buffer with protein carrier and preservative
Van Gogh Yellow Diluent (PD902)
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):
ISO
9001&13485
CERTIFIED
Protocol Recommendations (intelliPATH and manual use) Cont'd:
Protein Block (Optional): Incubate for 5-10 minutes at RT with Biocare's Background
Punisher.
Primary Antibody: Incubate for 30 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:
Counterstain with hematoxylin. Rinse with deionized water. Apply Tacha's Bluing
Solution for 1 minute. Rinse with deionized water.
intelliPATH™ Automated Slide Stainer:
IP016 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):
OAI016 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: LCA
Protocol Template (Description): Ms HRP Template 1
Dewaxing (DS Option): DS2
Antigen Retrieval (AR Option): AR1, high pH; 101°C
Reagent Name, Time, Temp.: LCA 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. Use TBS for washing steps. A standard PBS diluent (pH 7.2-7.4) is not
recommended for this antibody.
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.
Peroxide Block: Block for 5 minutes with Biocare's Peroxidazed 1.
Quality Control:
Pretreatment: Perform heat retrieval using Biocare's Reveal Decloaker. Refer to the
Reveal Decloaker product data sheet for specific instructions.
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
Page 1 of 2
Leukocyte Common Antigen (LCA) Cocktail
Concentrated and Prediluted Cocktail Antibody
Control Number: 901-016-010716
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) (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
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 SDS is available upon request and is located at http://biocare.net.
ISO
9001&13485
CERTIFIED
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. Muzaffar S, et al. Immunophenotypic analysis of non-Hodgkin's lymphoma. JPMA
J Pak Med Assoc. 1997 Apr; 47(4):106-9.
2. Michels S, et al. Immunostaining for leukocyte common antigen using an
Amplified avidin-biotin-peroxidase complex method and paraffin sections. A study of
735 hematopoietic and nonhematopoietic human neoplasms. Arch Pathol Lab Med.
1987 Nov; 111(11):1035-9.
3. Kurtin PJ, Pinkus GS. Leukocyte common antigen-a diagnostic discriminant
between hematopoietic and nonhematopoietic neoplasms in paraffin sections using
monoclonal antibodies: correlation with immunologic studies and ultrastructural
localization. Hum Pathol. 1985 Apr; 16(4):353-65.
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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