Mouse anti-Human Leucocyte Common Antigen (LCA) (unconjugated), Clone 2B11+PD7-26, IgG1, Purified

Concentrated Antibody for Immunohistochemistry

SKU:
K005
Category
Quantity
0, 5 ml
Buy at BIOZOL

Background

CD45 is a transmembrane glycoprotein expressed on most nucleated cells of haematopoietic origin. CD45, encoded by a single gene mapped to chromosome 1, has various isoforms based on differential splicing of exons 4, 5 and 6. On human leucocytes, five different isoforms of CD45, named ABC, AB, BC, B and 0, have been identified. These isoforms are recognized by CD45RA, CD45RB, CD45RC and CD45R0 antibodies. The isoforms range in Mr from 180 000 to 220 000. All the CD45 isoforms share the same intracellular segment, which has been shown to have tyrosine phosphatase activity. Various leucocytes express characteristic CD45 isoforms, thus T cells express CD45 isoforms corresponding to
their development and activation, B cells predominantly express the ABC isoform, and monocytes and dendritic cells predominantly express the B and 0 isoforms. Granulocytes principally express only the B and 0 isoforms (4).

Volume/Quantity

0,5 ml

Clone Number

2B11 + PD7/26

Preparation

Cell culture supernatant dialysed against 0.05 mol/L Tris/HCl, pH 7.2

Preservatives Stabilisers

15 mM Sodium Azide (NaN3)

Immunogen

2B11: Isolated neoplastic cells from a case of T-cell lymphoma/leukaemia (1).
PD7/26: Human peripheral blood lymphocytes maintained in T-cell growth factor (1).

Isotype

IgG1 (Mouse)

Specificity

Anti-CD45 is a mixture of two monoclonal antibodies, clones 2B11 and PD7/26, directed against different epitopes. Clone 2B11 was clustered as anti-CD45 at the Third International Workshop and Conference on Human Leucocyte Differentiation Antigens, held in Oxford in 1986 and reacts with all the known isotypes of the CD45 family (5). Clone PD7/26 was clustered as anti-CD45RB at the Fifth International Workshop and Conference on Human Leucocyte Differentiation Antigens, held in Boston in 1993 (6).
The antibody labels CD45 in both normal and neoplastic cells, and is a useful tool for identifying tumour cells of lymphoid origin (1-3). Differential identification is aided by the results from a panel of antibodies. Interpretation must be made within the context of the patient’s clinical history and other diagnostic tests by a qualified pathologist.

Fusion Partners

Spleen cells from an immunised BALB/c mouse were fused with cells of the mouse NSI myeloma cell line.

Suggested Working Dilution

FlowCytometry Not tested
Immunohistology-frozen Yes 1/50 – 1/100
Immunohistology-paraffin Yes 1/50 – 1/100
Immunohistology-resin Not tested
ELISA Not tested
Immunoprecipitation Not tested
Western Blotting Not tested
Radioimmunoassay Not tested

Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.

Immunohistology

Pre-treatment

Pretreatment of tissues with heat-induced epitope retrieval is recommended.

Positive Control Tissue

Spleen, thymus and lymph node

Recommended Secondary Reagents

F(ab’)2 rabbit anti-mouse IgG HRP conjugate – (LINARIS CatNo LST0013B)
ABC-Kit Mouse IgG-POD Labelling (LINARIS CatNo EDP4002)
DAB-Substrate for POD (LINARIS CatNo E108)
ABC-Kit Mouse IgG AP Labelling (LINARIS CatNo EDA5002)
BCIP/NBT Substrate for AP (LINARIS CatNo ESA5400)

Recommended Negative Controls

Mouse IgG1 Negative Control (LINARIS CatNo ITC0928)

Storage Conditions

Store at 2-8°C.
Should this product contain a precipitate we recommend microcentrifugation before use.

Shelf Life

12 months from date of despatch.

Warnke RA, Gatter KC, Falini B, Hildreth P, Woolston R-E, Pulford K, et al. Diagnosis of human lymphoma with monoclonal antileukocyte antibodies. N Engl J Med 1983;309:1275-81.

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;16:353.

Michie SA, Spagnolo DV, Dunn KA, Warnke RA, Rouse RV. A panel approach to the evaluation of the sensitivity and specificity of antibodies for the diagnosis of routinely processed histologically undifferentiated human neoplasms. Am J Clin Pathol 1987;88:457-62.

Sewell WA, Cooley MA, Hegen M. NL6. CD45 workshop panel report. In: Kishimoto T, Kikutani H, von dem Borne AEG, Goyert SM, Mason DY, Miyasaka M, et al., editors. Leucocyte typing VI. White cell differentiation antigens. Proceedings of the 6th International Workshop and Conference; 1996 Nov 10-14; Kobe, Japan. New York, London: Garland Publishing Inc.; 1997. p. 499-502.

Cobbold S, Hale G, Waldmann H. Non-lineage, LFA-1, and leucocyte common antigens: new and previously defined clusters. In: McMichael AJ, Beverley PCL, Cobbold S, Crumpton MJ, Gilks W, Gotch FM, et al., editors. Leukocyte typing III. White cell differentiation antigens. Proceedings of the 3rd International Workshop and Conference; 1986 Sep 21-26; Oxford, England. Oxford, New York, Tokyo: Oxford University Press; 1987. p. 788-803.

Morimoto C. T18. CD45 cluster report. In: Schlossman SF, Boumsell L, Gilks W, Harlan JM, Kishimoto T, Morimoto C, et al., editors. Leukocyte typing V. White cell differentiation antigens. Proceedings of the 5th International Workshop and Conference; 1993 Nov 3-7; Boston, USA. Oxford, New York, Tokyo: Oxford University Press; 1995. p. 386-9.

Herman GE and Elfont E. Aberrant CD45 (leukocyte common antigen) staining of non-malignant breast lesions in zinc formalin fixed tissue. J Histotechnol 1993;16:151-3.

Hall PA, d’Ardenne AJ, Stansfeld AG. Paraffin section immunohistochemistry. I. Non-Hodgkin’s lymphoma. Histopathol 1988;13:149- 60.

Specificity

Leucocyte Common Antigen (LCA)
Species Reactivity

Human
Host / Source

Mouse
Isotype

IgG1
Application

IHC-F, IHC-P
Clone

2B11+PD7-26
Antigen

Human Leucocyte Common Antigen (LCA)
Quantity

0, 5 ml
Format

Purified
Storage Temperature

2-8 °C
Shipping Temperature

2-8 °C

Documents

pdf Datasheet