فلوسایتومتری

Name: Flow Cytometry Antibody CD8-APC, Clone 17D8

Ø  Antibody CD8-APC is a monoclonal antibody (mAb) labelled with allophycocyanin (APC) designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD8 antigen-expressing cells.

SUMMARY AND EXPLANATION

FC is a powerful tool in analytical and quantitative characterization of cells which provides rapid and multiparametric analysis of heterogeneous cell populations on a cell-by-cell basis. FC is performed on cell suspension after incubating it with fluorescent-labelled antibodies directed against specific cellular proteins. Positive cells relative fluorescence intensity indicates the amount of antibody bonded to specific cell sites providing information about antigen expression. Human lymphocytes may be classified in three main populations according to their biological function and their cell surface antigen expression (T lymphocytes, B lymphocytes and natural killer (NK) cells). T lymphocytes (CD3+), the precursors of which originate in the bone marrow and then migrate and mature in the thymus, can be subdivided as well in functionally different populations. The most clearly defined of these are helper/inducer T cells (CD3+CD4+) and suppressor/cytotoxic T cells (CD3+CD8+). T cells produce no antibodies and are the mediators of cell immunity. CYT-8AP mAb recognizes CD8 antigen present on suppressor/cytotoxic T cells (CD3+CD8+) and can therefore be used in lymphocytes immunophenotyping. These studies are widely applied when monitoring post-transplant patient’s immunologic status and when characterizing and following-up immunodeficiencies, autoimmune diseases, leukaemia, etc.

REAGENT COMPOSITION

The purified monoclonal CD8 antibody conjugated with allophycocyanin (APC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

Clone: 17D8.

Isotype: IgG1.

Amount per vial: 200 tests (5 µl mAb per determination).

Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD9-Pacific Blue™, Clone MEM-61

Antibody CD9-Pacific Blue^TM is a monoclonal antibody (mAb) labelled with Pacific Blue^TM designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD9 antigen by flow cytometry (FC). This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

CD9 is a 24 kDa transmembrane protein also known as tetraspanin, MRP-1 and DRAP-24. It is expressed on platelets, monocytes, pre-B lymphocytes, granulocytes, activated T lymphocytes, endothelial cells and epithelial cells. Because of its aberrant expression in B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) cells, CD9 has been selected to contribute to the separation of BCP-ALL cells from normal/reactive BCP cells.

REAGENT COMPOSITION

Purified monoclonal CD9 antibody antibody conjugated with Pacific BlueTM, supplied in phosphate buffered saline with 0,09% sodium azide.

 Clone: MEM-61.

 Isotype: Mouse / IgG1.

Amount per vial: 100 tests (4 µL mAb per determination).

 Reagents are not considered sterile.

Name: Flow Cytometry Antibody CD10-APC, Clone HI10a

• CD10-APC is designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD10 antigen-expressing cells. This reagent must be used by FC qualified personal.

SUMMARY AND EXPLANATION

CYT-10AP mAb recognizes CD10 antigen, a 100 KDa glycoprotein belonging to a family of type II transmembrane metalloproteases. This molecule is expressed on immature T and B precursor cells but is lost as the cells reach maturation. However, it is re-expressed on proliferating B cells and mature neutrophils. CD10 antigen is a common acute lymphoblastic leukaemia associated antigen (CALLA) as it is found on lymphocytes from acute B-lymphoid leukaemia samples. It is also present on a wide variety of normal and neoplastic cell types including renal epithelium, fibroblasts, granulocytes, and some T-cell leukaemia, lymphoma, melanoma, and glioma cell lines. This antibody can therefore be used in the characterization of non-T (common) acute lymphoblastic leukemias (8, 9) and in the studies of early stages of hematopoietic differentiation.

REAGENT COMPOSITION

The purified monoclonal CD10 antibody conjugated with allophycocyanin (APC) is supplied in phosphate-buffered saline (PBS) containing 0,09% (m/v) sodium azide.

Clone: HI10a.

 Isotype: IgG1.

Amount per 0.5 mL vial: 100 tests (5 µL mAb per determination).

Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD10-APC-C750, Clone HI10a

• CD10-APC-C750^TM is designed for flow cytometry (CF) use as a direct immunofluorescence reagent in the identification and enumeration of CD10 antigen-expressing cells. This reagent must be used by FC qualified personnel.

SUMMARY AND EXPLANATION

CYT-10AC750 mAb recognizes CD10 antigen, a 100 KDa glycoprotein belonging to a family of type II transmembrane metalloproteases. This molecule is expressed on immature T and B precursor cells but is lost as the cells reach maturation. However, it is re-expressed on proliferating B cells and mature neutrophils. CD10 antigen is a common acute lymphoblastic leukaemia associated antigen (CALLA) as it is found on lymphocytes from acute B-lymphoid leukaemia samples. It is also present on a wide variety of normal and neoplastic cell types including renal epithelium, fibroblasts, granulocytes, and some T-cell leukaemia, lymphoma, melanoma, and glioma cell lines. This antibody can therefore be used in the characterization of non-T (common) acute lymphoblastic leukemias and in the studies of early stages of hematopoietic differentiation. APC-C750^TM is a tandem dye with a maximum emission peak at 779 nm, which grants bright signal, low unspecific noise and high photostability. When excited by light from a red laser, the APC fluorochrome can transfer energy to C750 molecule, which then emits at a longer wavelength. It is recommended to use a 780/60 nm longpass filter along with a red sensitive detector to use in conjunction antibodies conjugated with APC and APC-C750^TM.

REAGENT COMPOSITION

The purified monoclonal CD10 antibody conjugated with the tandem allophycocyanine-C750 (APC-C750TM) is supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: HI10a.
• Isotype: IgG1.
• Amount per 0,15 mL vial: 50 tests (3 µL mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD14-APC, Clone 47-3D6

• Antibody CD14 is a monoclonal antibody (mAb) labelled with different fluorochromes (see table) and designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD14 antigen expressing cells. This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

Leukocytes are named according to structure as being either granulocytes or agranulocytes, and according to the function as either phagocytes or immunocyte. The granulocytes, which include neutrophils, basophils and eosinophils, are all phagocytes. Of the agranulocytes, the monocytes and macrophages are phagocytes whereas the lymphocytes are immunocytes. Monocytes ingest dead or damaged cells and help defend against many infectious organisms. Anti-CD14 mAb recognizes CD14 antigen present on monocyte/macrophage cells. The reagent is important for excluding CD14+ monocytes in relation to T lymphocytes subsets analysis. In FC, anti-CD14 is considered essential for the initial evaluation of acute myeloid leukaemias together with a panel of other antibodies.

REAGENT COMPOSITION

The purified monoclonal CD14 antibody conjugated with different fluorochromes (see table above) is supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: 47-3D6.
• Isotype: Mouse / IgG1.
• Amount per 1 ml vial: 200 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD3-PE- Cyanine 5, Clone UCHT-1

• Antibody CD3 reagent is a monoclonal antibody (mAb) conjugated with different fluorochromes (see table above) and designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD3 antigen by flow cytometry. This reagent must be used by flow cytometry qualified personnel.

SUMMARY AND EXPLANATION

The anti-CD recognizes the CD3 antigen present in T cells and can therefore be used in the characterization studies for immunophenotyping of lymphocytes. These studies are widely applied for monitoring of the immunologic status of posttransplant patients and in the characterization and follow-up of immunodeficiencies, autoimmune diseases, leukemia etc. The T lymphocyte (CD3+) count is generally expressed as a percentage of the total amount of lymphocytes or leucocytes present in the sample which can itself be determined by flow cytometry based on its typical pattern of FSC/SSC (size/granularity or complexity). Because each flow cytometer has different operating characteristics each laboratory must determine its optimal operating procedure.

REAGENT COMPOSITION

Purified monoclonal CD3 antibody conjugated with different fluorochromes (see table above) is supplied in phosphatebuffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: UCHT-1
• Isotypes: Mouse / IgG1
• Purification: Affinity chromatography
• Amount per vial: 50 or 200 tests (3 or 5 µl mAb to 106 cells).
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody B2-microglobulin-PerCP-Cyanine5.5, Clone B2-1

• Antibody β2-Micro-PerCP-Cyanine5.5 is a monoclonal antibody (mAb) labelled with Peridin chlorophyll protein-Cyanine 5.5 (PerCPCyanine5.5), designed for Flow Cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of β2-Micro antigen-expressing cells. This reagent must be used by FC qualified personal.

SUMMARY AND EXPLANATION

β2-Micro-PerCP-Cyanine5.5 mAb recognizes B2-Micro antigen, a 12-kilodalton (kDa) protein with homology to the constant region of immunoglobulin molecules. The β2-Micro antigen is found on the surface of most nucleated cells. The β2-Micro antigen heterodimerizes with a polymorphic heavy chain to form the major histocompatibility complex (MHC) class I molecule.

REAGENT COMPOSITION

The purified monoclonal β2-Micro antibody conjugated with the tandem PerCP-Cyanine5.5 is supplied in phosphatebuffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: B2-1.
• Isotype: Mouse / IgG2a.
• Amount per vial: 50 tests (3 µl mAb per determination).
• Reagent is considered non-sterile.