Monoclonal
Antibodies
Detecting
Human
Antigens
RESEARCH
APPLICATIONS
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BD Oncomark™
Lambda/Kappa/CD5/CD10/CD19/CD45
Catalog No. 338428
50 Tests
20 µL/test
Research applications include:
•
Studies of B-cell biology
DESCRIPTION
Specificity
Anti-Lambda is specific for lambda light chains of human immunoglobulins.1
Anti-Kappa is specific for kappa light chains of human immunoglobulins.
CD5 recognizes a human T-lymphocyte antigen, with a molecular weight of
67 kilodaltons (kDa).2
CD10 (Anti-CALLA) recognizes a human common acute lymphoblastic leukemia
antigen (CALLA), with a molecular weight of 100 kDa.3,4 The CD10 antigen is
identical to human membrane–associated neutral endopeptidase (NEP; EC 3.3.24.11),
also known as enkephalinase.5
CD19 (SJ25C1) recognizes a 90-kDa antigen that is present on human
B lymphocytes.6,7
CD45 (Anti–HLe-1) recognizes human leucocyte antigens, with a molecular weight of
180 to 220 kDa, that are members of the T200 family.8
Antigen distribution
Immunoglobulins bearing lambda light chains are present on approximately 40% of
normal B lymphocytes and on Igλ+ neoplastic cells.9-15 In serum, Anti-Lambda reacts
with immunoglobulins bearing lambda light chains as well as free lambda light chains.
Immunoglobulins bearing kappa light chains are present on approximately 60% of
normal B lymphocytes and on Igκ+ neoplastic cells.9-15 In serum, Anti-Kappa reacts
with immunoglobulins bearing kappa light chains as well as free kappa light chains.
The CD5 antigen is present on approximately 70% of normal peripheral blood
lymphocytes and on virtually all T lymphocytes in thymus and peripheral blood.16-18
The CD5 antibody reacts with most cells in T-lymphocyte areas of spleen and lymph
node and with many T-cell leukemias and lymphomas.19-21 It also reacts with a distinct
subset of normal B lymphocytes,22 occasional cells in B-lymphocyte areas of spleen and
lymph node,19 and most Ig+ B–chronic lymphoblastic leukemia (CLL) cells.21-25 Some
lymphomas also express the CD5 antigen.20
The CD10 antigen is found on lymphocytes from acute B-lymphoid leukemia
samples.26,27 The antigen is also present on a wide variety of normal and neoplastic cell
types including renal epithelium, fibroblasts, granulocytes, and some lymphoma,
melanoma, and glioma cell lines.5
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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BD Biosciences
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San Jose, CA 95131 USA
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01/2015
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The CD19 antigen is present on approximately 7% to 23% of human peripheral blood
lymphocytes28 and on splenocytes.29 The CD19 antigen is present on human
B lymphocytes at most stages of maturation.7,30 CD19 does not react with resting or
activated T lymphocytes, granulocytes, or monocytes.7,30
The CD45 antigen is present on all human leucocytes, including lymphocytes,
monocytes, granulocytes, eosinophils, and basophils in peripheral blood and has a role
in signal transduction, modifying signals from other surface molecules.8 The CD45
antibody has been reported to react weakly with mature circulating erythrocytes and
platelets.8,31
Clones
Anti-Lambda, clone 1-155-2,* is derived from hybridization of P3-X63-Ag8.653 mouse
myeloma cells with cells from BALB C/J mice immunized with human IgA1-λ myeloma
protein.
Anti-Kappa, clone TB28-2,* is derived from hybridization of P3-X63-Ag8.653 mouse
myeloma cells with cells from CB6 (BC57b x BALB/c) mice immunized with human
IgG-κ myeloma protein.
CD5, clone L17F12,16 is derived from hybridization of NS-1/Ag4 mouse myeloma cells
with spleen cells from BALB/c mice immunized with human T-acute lymphoblastic
leukemia cells.
CD10, clone HI10a,4 is derived from the hybridization of P3-63-Ag8.653 mouse
myeloma cells with spleen cells of BALB/c mice immunized with blasts from a patient
with acute CALLA leukemia.
CD19, clone SJ25C1,7 is derived from the hybridization of Sp2/0 mouse cells with
spleen cells from BALB/c mice immunized with NALM1 + NALM16 cells.
CD45, clone 2D1,8 is derived from hybridization of NS-1 mouse myeloma cells with
spleen cells from BALB/c mice immunized with human peripheral blood mononuclear
cells (PBMCs).
Composition
Anti-Kappa, CD19, and CD45 are each composed of mouse IgG1 heavy chains and
kappa light chains.
Anti-Lambda is composed of mouse IgG1 heavy chains and lambda light chains.
CD5 and CD10 are each composed of mouse IgG2a heavy chains and kappa light
chains.
This BD Oncomark™ reagent is supplied as a combination of Anti-Lambda FITC,
Anti-Kappa PE, CD5 PerCP-Cy™5.5, CD10 PE-Cy™7, CD19 APC, and CD45
APC-Cy7 in 1 mL of phosphate-buffered saline (PBS) containing gelatin and 0.1%
sodium azide.
PROCEDURE
Visit our website (bdbiosciences.com) or contact your local BD representative for the
lyse/wash protocol for direct immunofluorescence.
To avoid serum interference when using this reagent:
1. Prewash the whole blood sample using at least 25 volumes of excess 1X PBS with
0.1% sodium azide (For example, 48 mL of 1X PBS with sodium azide per 2 mL of
whole blood to be washed) and mix well.
2. Pellet cells by centrifugation.
3. Resuspend in 1X PBS with 0.1% sodium azide to the original volume.
NOTE Spectral overlap values for PE-Cy7 and APC-Cy7 conjugates can vary from lot
to lot. It is important to check these values on a known sample when using a new lot of
reagents.
*
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This clone has not been submitted to any previous workshop on human leucocyte differentiation antigens.
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CAUTION Some APC-Cy7 conjugates, and to a lesser extent PE-Cy7 and APC-H7
conjugates, show changes in their emission spectra with prolonged exposure to
paraformaldehyde or light. For overnight storage of stained cells, wash and resuspend
in buffer without paraformaldehyde after 1 hour of fixation.We recommend that you
analyze fixed samples within four hours.
REPRESENTATIVE DATA
Performed on whole blood stained and lysed using BD FACS™ lysing solution (Cat.
No. 349202).
SSC-A
250
250
Figure 1 Representative data analyzed with a BD FACS™ brand flow cytometer
SSC-A
granulocytes
50
50
monocytes
lymphocytes
250
FSC-A
102
105
CD10 PE-Cy7
105
SSC-A
SSC-A
50
50
CD19+
102
CD19 APC
105
CD5+CD19–
CD5+CD19+
Q4
Q2
Anti-Kappa+
Q3
Anti-Lambda+
102
Q3
Anti-Kappa PE 105
102
102 CD5 PerCP-Cy5.5 105
CD45 APC-Cy7
250
250
50
102
CD19 APC
105
102
CD20 APC-Cy7
105
HANDLING AND
STORAGE
Store vials at 2°C–8°C. Conjugated forms should not be frozen. Protect from exposure
to light. Each reagent is stable until the expiration date shown on the bottle label when
stored as directed.
WARNING
All biological specimens and materials coming in contact with them are considered
biohazards. Handle as if capable of transmitting infection32,33 and dispose of with
proper precautions in accordance with federal, state, and local regulations. Never
pipette by mouth. Wear suitable protective clothing, eyewear, and gloves.
CHARACTERIZATION
To ensure consistently high-quality reagents, each lot of antibody is tested for
conformance with characteristics of a standard reagent. Representative flow cytometric
data is included in this data sheet.
Page 3
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WARRANTY
Unless otherwise indicated in any applicable BD general conditions of sale for non-US
customers, the following warranty applies to the purchase of these products.
THE PRODUCTS SOLD HEREUNDER ARE WARRANTED ONLY TO CONFORM TO THE QUANTITY AND CONTENTS
STATED ON THE LABEL OR IN THE PRODUCT LABELING AT THE TIME OF DELIVERY TO THE CUSTOMER. BD
DISCLAIMS HEREBY ALL OTHER WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING WARRANTIES OF
MERCHANTABILITY AND FITNESS FOR ANY PARTICULAR PURPOSE AND NONINFRINGEMENT. BD’S SOLE LIABILITY
IS LIMITED TO EITHER REPLACEMENT OF THE PRODUCTS OR REFUND OF THE PURCHASE PRICE. BD IS NOT LIABLE
FOR PROPERTY DAMAGE OR ANY INCIDENTAL OR CONSEQUENTIAL DAMAGES, INCLUDING PERSONAL INJURY,
OR ECONOMIC LOSS, CAUSED BY THE PRODUCT.
REFERENCES
1.
Kubagawa H, Gaithings WE, Levitt D, Kearney JF, Cooper MD. Immunoglobulin isotype expression of
normal pre-B cells as determined by immunofluorescence. J Clin Immunol. 1982;2:264.
2.
Knowles RW. Immunochemical analysis of the T-cell–specific antigens. In: Reinherz EL, Haynes BF,
Nadler LM, Bernstein ID, eds. Leukocyte Typing II: Human T Lymphocytes. New York, NY: SpringerVerlag; 1986;1:259-288.
3.
Greaves MF. Monoclonal antibodies as probes for leukemic heterogeneity and hematopoietic
differentiation. In: Knapp W, ed. Leukemia Markers. New York, NY: Academic Press; 1981:19.
4.
Zola H. CD10 Workshop Panel report. In: Schlossman SF, Boumsell L, Gilks W, et al, eds. Leucocyte
Typing V: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1995:505-507.
5.
Letarte M, Vera S, Tran R, et al. Common acute lymphocytic leukemia antigen is identical to neutral
endopeptidase. J Exp Med. 1988;168:1247-1253.
6.
Moldenhauer G, Dörken B, Schwartz R, Pezzutto A, Knops J, Hammerling GJ. Analysis of ten
B lymphocyte-specific workshop monoclonal antibodies. In: Reinherz EL, Haynes BF, Nadler LM,
Bernstein ID, eds. Leukocyte Typing II: Human B Lymphocytes. New York, NY: Springer-Verlag;
1986;2:61-67.
7.
Nadler LM. B Cell/Leukemia Panel Workshop: summary and comments. In: Reinherz EL, Haynes BF,
Nadler LM, Bernstein ID, eds. Leukocyte Typing II: Human B Lymphocytes. New York, NY: SpringerVerlag; 1986;2:3-43.
8.
Schwinzer R. Cluster report: CD45/CD45R. In: Knapp W, Dörken B, Gilks WR, et al, eds. Leucocyte
Typing IV: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989:628-634.
9.
Chung J, Gong G, Huh J, Khang SK, Ro JY. Flow cytometric immunophenotyping in fine-needle
aspiration of lymph nodes. J Korean Med Sci. 1999;14:393-400.
10. Zardawi IM, Jain S, Bennett G. Flow-cytometric algorithm on fine-needle aspirates for the clinical
workup of patients with lymphadenopathy. Diagn Cytopathol. 1998;19:274-278.
11. Davidson B, Risberg B, Berner A, Smeland EB, Torlakovic E. Evaluation of lymphoid cell populations in
cytology specimens using flow cytometry and polymerase chain reaction. Diagn Mol Pathol. 1999;8:183-188.
12. Braylan RC, Benson NA, Iturraspe J. Analysis of lymphomas by flow cytometry: current and emerging
strategies. Ann NY Acad Sci. 1993;677:364-378.
13. Johnson A, Cavallin-Stahl E, Akerman M. Flow cytometric light chain analysis of peripheral blood
lymphocytes in patients with non-Hodgkin's lymphoma. Br J Cancer. 1985;52(2):159-165.
14. Letwin BW, Wallace PK, Muirhead KA, Hensler GL, Kashatus WH, Horan PK. An improved clonal
excess assay using flow cytometry and B-cell gating. Blood. 1990;75:1178-1185.
15. Geary WA, Frierson HF, Innes DJ, Normansell DE. Quantitative criteria for clonality in the diagnosis of
B-cell non-Hodgkin's lymphoma by flow cytometry. Mod Pathol. 1993;6:155-161.
16. Engleman EG, Warnke R, Fox RI, Dilley J, Benike CJ, Levy R. Studies of a human T lymphocyte antigen
recognized by a monoclonal antibody. Proc Natl Acad Sci USA. 1981;78:1791-1795.
17. Ledbetter JA, Evans RL, Lipinski M, Cunningham-Rundles C, Good RA, Herzenberg LA. Evolutionary
conservation of surface molecules that distinguish T-lymphocyte helper/inducer and
T cytotoxic/suppressor subpopulations in mouse and man. J Exp Med. 1981;153:310-323.
18. Ledbetter JA, Frankel AE, Herzenberg LA. Human Leu T-cell differentiation antigens: quantitative expression on
normal lymphoid cells and cell lines. In: Hämmerling G, Hämmerling U, Kearney J, eds. Monoclonal Antibodies
and T Cell Hybridomas: Perspectives and Technical Notes. New York, NY: Elsevier/North Holland;
1981:16-22.
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19. Warnke RA, Levy R. Detection of T and B antigens with hybridoma monoclonal antibodies: a biotinavidin-horseradish peroxidase method. J Histochem Cytochem. 1980;28:771-776.
20. Warnke R, Miller R, Grogan T, Pederson M, Dilley J, Levy R. Immunologic phenotype in 30 patients
with diffuse large-cell lymphoma. N Eng J Med. 1980;303:293-300.
21. Zipf RF, Fox R, Dilley J, Levy R. Definition of the high risk ALL patient by immunologic phenotyping
with monoclonal antibodies. Cancer Res. 1981;41:4786.
22. Gadol N, Ault KA. Phenotypic and functional characterization of human Leu-1 (CD5) B cells. Immunol
Rev. 1986;93:23.
23. Royston I, Majda JA, Baird SM, Meserve BL, Griffiths JC. Human T-cell antigens defined by monoclonal
antibodies: the 65,000-dalton antigen of T cells (T65) is also found on chronic lymphocytic leukemia
cells bearing surface immunoglobulin. J Immunol. 1980;125:725.
24. Dono M, Cerruti G, Zupo S. The CD5+ B-cell. Int J Biochem Cell Biol. 2004;36:2105-2111.
25. D'Arena G, Di Renzo N, Brugiatelli M, Vigliotti ML, Keating MJ. Biological and clinical heterogeneity of
B-cell chronic lymphocytic leukemia. Leuk Lymphoma. 2003;44:223-228.
26. Le Bien TW, McCormack RT. The common acute lymphoblastic leukemia antigen (CD10): emancipation
from a functional enigma. Blood. 1989;73:625-635.
27. Morabito F, Mangiola M, Rapezzi D, et al. Expression of CD10 by B-chronic lymphocytic leukemia cells
undergoing apoptosis in vivo and in vitro. Haematologica. 2003;88:864-873.
28. Reichert T, DeBruyère M, Deneys V, et al. Lymphocyte subset reference ranges in adult Caucasians. Clin
Immunol Immunopath. 1991;60:190-208.
29. Tedder T, Zhou L-J, Engel P. The CD19/CD21 signal transduction complex of B lymphocytes. Immunol
Today. 1994;15:437-442.
30. Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. II.
Normal B-lymphocyte development. Blood. 1987;70:1316-1324.
31. Jackson A. Basic phenotyping of lymphocytes: selection and testing of reagents and interpretation of
data. Clin Immunol Newslett. 1990;10:43-55.
32. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline —
Third Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2005. CLSI document M29-A3.
33. Centers for Disease Control. Perspectives in disease prevention and health promotion update: universal
precautions for prevention of transmission of human immunodeficiency virus, hepatitis B virus, and
other bloodborne pathogens in health-care settings. MMWR. 1988;37:377-388.
PATENTS AND
TRADEMARKS
APC-Cy7: US Patent Number 5,714,386
Cy™ is a trademark of GE Healthcare. This product is subject to proprietary rights of
GE Healthcare and Carnegie Mellon University, and is made and sold under license
from GE Healthcare. This product is licensed for sale only for research. It is not
licensed for any other use. If you require a commercial license to use this product and
do not have one, return this material, unopened, to BD Biosciences, 2350 Qume Drive,
San Jose, CA 95131, and any money paid for the material will be refunded.
BD, BD Logo and all other trademarks are property of Becton, Dickinson and
Company. © 2015 BD
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