فلوسایتومتری

Name: Flow Cytometry Antibody CD45-PE-Cyanine5, Clone HI30

• Reagent CD45 is a monoclonal antibody (mAb) labelled with different fluorochromes (see table) and designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD45 antigen by Flow Cytometry (FC). This reagent must be used by flow cytometry qualified personnel.

        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. The granulocytes have granules in their cytoplasm, which contain enzymes which are capable of killing microorganisms and destroying debris ingested by the process of phagocytosis. The anti-CD45 mAb reacts with the leukocyte common antigen (LCA) complex, which is present on lymphocytes, monocytes, granulocytes, eosinophils, basophils and their progenitors. This reagent can be used in the characterization studies for immunophenotyping of leukocytes, which are widely applied in the characterization and follow-up of immunodeficiencies, autoimmune diseases, leukemias, etc.

REAGENT COMPOSITION

Purified monoclonal CD45 antibody conjugated with different fluorochromes (see table above) and supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: HI30
• Isotype: Mouse / IgG1
• Purification: Affinity chromatography
• Amount per vial: 200/50 tests (5 or 3 µl mAb to 106 cells (see table above))
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody CD45-PerCP-Cyanine5.5 (HI30), Clone HI30

• Antibody Reagent CD45 is a monoclonal antibody (mAb) labelled with different fluorochromes (see table) and designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD45 antigen by Flow Cytometry (FC). This reagent must be used by flow cytometry qualified personnel.

       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. The granulocytes have granules in their cytoplasm, which contain enzymes which are capable of killing microorganisms and destroying debris ingested by the process of phagocytosis. The anti-CD45 mAb reacts with the leukocyte common antigen (LCA) complex, which is present on lymphocytes, monocytes, granulocytes, eosinophils, basophils and their progenitors. This reagent can be used in the characterization studies for immunophenotyping of leukocytes, which are widely applied in the characterization and follow-up of immunodeficiencies, autoimmune diseases, leukemias, etc.

REAGENT COMPOSITION

Purified monoclonal CD45 antibody conjugated with different fluorochromes (see table above) and supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: HI30
• Isotype: Mouse / IgG1
• Purification: Affinity chromatography
• Amount per vial: 200/50 tests (5 or 3 µl mAb to 106 cells (see table above))
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody CD49d-FITC, Clone 44H6

• Antibody CD49d-FITC is a monoclonal antibody (mAb) labelled with fluorescein isothiocyanate (FITC) designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD49d 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. Flow cytometry is performed on cell suspension after incubating it with fluorescentlabelled 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. During inflammatory processes, leucocytes interact with extracellular matrix and vascular endothelium through membrane receptors from the integrin family. These interactions are transient and allow leucocytes to find antigens, respond to inflammatory stimuli, extravasate or adhere firmly to the endothelium to perform immune functions .CD49d is an integrin subunit (α4 chain) that pairs with two other β integrin subunits: CD29 (β1 chain) to form a very late activation antigen 4 (VLA-4) or in lesser proportion with β7 chain to form the α4β7 dimer. VLA-4 is expressed at significant levels on all circulating leukocytes except mature neutrophils; α4β7 is expressed on a subset of T and B cells, natural killer cells, and eosinophils. CD49d has a role in leucocyte recruitment in a variety of inflammatory and immune disorders. CYT-49dF mAb is useful for studies of leucocyte integrin molecules, cell adhesion, T-lymphocyte cytokine production, T-cell activation, haematogenous tumour metastasis, expression of integrin α4 on melanoma and Burkitt lymphoma, etc.

REAGENT COMPOSITION

The purified monoclonal CD49d antibody conjugated with fluorescein isothiocyanate (FITC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

• Clone: 44H6.
• Isotype: IgG1.
• Amount per 1 ml vial: 200 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD59-FITC, Clone MEM-43

• Antibody CD59-FITC is a monoclonal antibody (mAb) labelled with fluorescein isothiocyanate (FITC) designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD59 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. Flow cytometry is performed on cell suspension after incubating it with fluorescentlabelled 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. Membrane-associated C-regulatory proteins are the major protection against Complement attack. CD59 (Protectin), a 19 kDa GPI-linked membrane glycoprotein is one of these regulatory proteins binding to the complement components C8 and C9 preventing the destruction of autologous cells. CD59 has a broadly distributed expression on all lymphocytes, monocytes, granulocytes, erythrocytes and in a variety of non-haematological cells. CD59 modulates T cell adhesion and plays a role in T cell activation. CD59 deficiency was already detected in clinical inflammatory disorders. Cells from patients with the haematological disorder paroxysmal nocturnal haemoglobinuria (PNH) are defective for GPI anchor synthesis and hence lack surface expression of CD59 and are highly susceptible to the lytic effects of Complement attack. For this reason CD59 together with CD55, another GPI-anchored protein, have been systematically used in diagnosis of PNH.

REAGENT COMPOSITION

The purified monoclonal CD59 Antibody conjugated with fluorescein isothiocyanate (FITC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

• Clone: MEM-43
• Isotype: IgG2a.
• Amount per 1 mL vial: 200 tests (5 µL mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD66b-FITC, Clone 80H3

• Antibody CD66b-FITC is a monoclonal antibody (mAb) labelled with fluorescein isothiocyanate (FITC) designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD66b 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. Flow cytometry is performed on cell suspension after incubating it with fluorescentlabelled 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. Haematopoietic cells give rise to all blood cell types from myeloid (e.g. monocytes, neutrophils, basophils and erythrocytes) and lymphoid (e.g. T, B and NK cells) lineages. Several markers have been routinely used to distinguish both lineages; an example is the CD66b which is used to identify mature granulocytes (neutrophils and eosinophils). CD66b is a GPI-linked protein member of the carcinoembryonic antigen (CEA)-like glycoprotein that was first described in colon cancer cells and is not expressed in normal peripheral B cells, T cells, monocytes and platelets. CD66b has an increase in its expression in granulocytes activated with soluble stimulators (calcium ionophore, phorbol myristate acetate, nFMLP). CD66 proteins are involved in several cellular processes (e.g. cell adhesion and activation). Eosinophils CD66b expression increases in patients with Helminth infection. Also, leukocytes of Chronic Myeloid Leukaemia (CML) present CD66b expression.

REAGENT COMPOSITION

The purified monoclonal CD66b Antibody conjugated with fluorescein isothiocyanate (FITC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

• Clone: 80H3.
• Isotype: IgG1.
• Amount per 1 ml vial: 200 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD81-FITC, Clone M38

• Antibody Anti human CD81-FITC is a monoclonal antibody (mAb) labelled with fluorescein isothiocyanate (FITC) designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD81 antigen by flow cytometry (FC). This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

The CD81 (TAPA-1) antigen is expressed on normal lymphocytes, monocytes and eosinophils, but not on platelets and neutrophils. This antigen may be present in some cases as multimolecular complexes, in association with other members of the TM4 superfamily (CD37, CD53), or on the surface of B cells, in association with CD19 and/or CD21 and/or MHC class II antigens. Most B lymphocytes, at all stages of cellular differentiation, express CD81 at relatively high levels. Because each flow cytometer has different operating characteristics each laboratory must determine its optimal operating procedure.

REAGENT COMPOSITION

Purified monoclonal CD81 antibody conjugated with fluorescein isothiocyanate (FITC), supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: M38.
• Isotype: Mouse / IgG1.
• Amount per vial: 100 tests (5 µL mAb per determination)
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody CD134-PE, Clone 134-1

• Antibody CD134-PE is a monoclonal antibody (mAb) labelled with R-phycoerythrin (PE) designed for use as a direct immunofluorescence reagent in the identification and enumeration of CD134 antigen-expressing cells. This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

The mAb, CYT-134PE, specifically binds to CD134 which is also known as OX40. CD134 is a type I integral membrane glycoprotein and member of the tumor necrosis factor/nerve growth factor receptor (TNFR/NGFR) family. CD134 is expressed on activated T lymphocytes, hematopoietic precursor cells and fibroblasts. CD134 plays roles in T-cell activation as well as the regulation of differentiation, proliferation or apoptosis of normal and malignant lymphoid cells

REAGENT COMPOSITION

The purified monoclonal CD134 antibody conjugated with R-phycoerythrin (PE) is supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: 134-1.
• Isotype: IgG1.
• Amount per vial: 100 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD138, Clone B-A38

• Antibody CD138 reagent is a monoclonal antibody (mAb) conjugated with different fluorochromes (see table) and designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of human CD138 antigen-expressing cells. This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

The mAb Anti-CD138 reacts with human CD138, syndecan-1, a 20-22 kDa protein that binds to interstitial extracellular matrix molecules. CD138 is predominantly expressed on epithelial cells and B lymphocytes at specific stages in their differentiation: precursor B cells in the bone marrow and antibody-secreting cells, including plasma cells but not mature peripheral B cell. The simultaneous assessment of the expression of CD38 and CD138 represents the best combination of markers for the identification of plasma cells.

REAGENT COMPOSITION

The purified mAb CD138 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: B-A38.
• Isotype: Mouse / IgG1.
• Reagent is considered non-sterile.