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Name: Flow Cytometry Antibody CD16+ CD56-PE, Clone 3G8, C5.9

• Antibody CD16+CD56-PE is a single-color direct immunofluorescence reagent for use in flow cytometry (FC) designed for the identification and enumeration of Natural Killer (CD3-CD16+CD56+) cells. This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

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 cells (NK). NK cells (CD16+CD56+) mediate cytotoxicity against certain tumors and virus-infected cells. NK-mediated cytotoxicity does not require class I or class II major histocompatibility complex (MHC) molecules to be present on the target cell. The CD16+CD56-PE reagent recognizes the antigens CD16 and CD56 present in NK cells. This reagent can be used in the characterization studies for immunophenotyping of lymphocytes. These studies are widely applied in the characterization and follow-up of acute and chronic lymphoproliferative disorders, immunodeficiencies, autoimmune diseases. The CD16 antigen is also detected on macrophages and neutrophils. The CD56 antigen is also detected on a subset of cytotoxic T lymphocytes that mediates non-MHC-restricted cytotoxicity.

REAGENT COMPOSITION

CD16+CD56-PE reagent is supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide. It contains the following mixture of mAb:

• Phycoerythrin (PE)-labeled CD16, clone 3G8, isotype IgG1
• Phycoerythrin (PE)-labeled CD56, clone C5.9, isotype IgG2b.

Purification: Affinity chromatography

Amount per 1 ml vial: 100 tests (10 ml reagent to 106 cells)

Reagents are not considered sterile.

Name: Flow Cytometry Antibody cyTCRβ-APC, Clone 8A3

• Antibody cyTCRβ-APC is a monoclonal antibody (mAb) labelled with allophycocyanine (APC) designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the β chain of the T-cell Receptor (TCR -βF1) complex by flow cytometry (FC).

SUMMARY AND EXPLANATION

FC is a powerful tool for the analytical and quantitative characterization of cells which provides rapid, quantitative and multiparametric analysis of heterogeneous cell populations on a cell-by-cell basis. FC is performed on cells in liquid suspension that have been incubated with fluorescently labeled antibodies directed against specific cellular proteins. The relative fluorescence intensity of the positive cells indicates the amount of antibody bound to specific binding sites on the cells, and therefore provides a relative measure of antigen expression. The 8A3 antibody is specific for human TCR β chain constant region which is expressed by αβ TCR-expressing thymocytes and peripheral T lymphocytes. It does not react with γδ TCR-bearing T cells. In the fetal and adult thymus, the TCR β-chain may form homodimers or pair with the pre-TCR α-chain on the surface of immature thymocytes before expression of the TCR α-chain. The 8A3 antibody has been cited in several research papers involving the study of lymphoma. It is most often used for tissue staining and has been used to stain both frozen sections and paraffin-embedded tissue but is also suitable for immunoprecipitation and FC.

REAGENT COMPOSITION

Purified monoclonal TCR -βF1 antibody conjugated with allophycocyanine (APC), supplied in phosphate buffered saline with ≤0,09% (m/v) sodium azide.

• Clone: 8A3.
• Isotype: Mouse / IgG1.
• Amount per vial: 100 tests (5 µl mAb per determination).
• 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.
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody CYT-MPOF3, Clone 2C7

• Antibody CYT-MPOF3 is a monoclonal antibody (mAb) anti human Myeloperoxidase labelled with fluorescein isothiocyanate (FITC) designed for use as a direct immunofluorescence reagent in the identification and enumeration of myelomonocytic cells by flow cytometry. This reagent must be used by flow cytometry qualified personal.

REAGENT COMPOSITION

Purified monoclonal cyMPO antibody conjugated with fluorescein isothiocyanate (FITC), supplied in phosphate buffered saline with ≤0,1 % (m/v) sodium Azide.

• Clone: 2C7
• Isotype: Mouse / IgG1
• Size: 100 tests (0,5 ml per vial) Usage: 5 µl/ test
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody Cytokeratin18-FITC, Clone Ks18.04

• Antibody Ks 18.04 represents an excellent marker to discriminate simple epithelia from those of different origin. Tumors specifically detected: all adenocarcinoma; mammary carcinoma, urinary bladder carcinoma, undifferentiated carcinoma, cervix carcinoma, hepatocellular carcinoma. Polypeptide reacting: Mr 45 000 polypeptide (human cytokeratin 18) of all simple type epithelium and basal cells of many squamous, nonepidermal epithelium.

Suitable for:

– Immunohistochemistry of frozen and paraffinembedded tissue and cytological material (Work dilution 1:10)

– Cell labeling for later analysis by flow cytometry – This reagent must be used by flow cytometry qualified personal.

REAGENT COMPOSITION

Purified mouse monoclonal antibody to cytokeratin 18-FITC conjugate, supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and ≤0.09% (m/v) sodium azide.

• Clone: Ks18.04
• Isotype: IgG1
• Purification: Protein A affinity chromatography
• Amount per vial: 0.25 ml

Name: Flow Cytometry Antibody HLA-B27-FITC, Clone HLA–ABC–m3

• HLA-B27-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 HLA-B27 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. Human Leukocyte Antigen (HLA) B27 is a class I surface antigen encoded by the B locus in the major histocompatibility complex (MHC) on chromosome 6 and presents antigenic peptides to T cells. HLA-B27 and HLAB7 antigens are expressed in 7% and 22% of individuals of Caucasian origin. This mixture of antibodies permits the characterization of HLA-B27 specificity in the HLA class I allotype in patients suffering from inflammatory disorders affecting the sacroiliac and intervertebral joints. This finding aids in the diagnosis of ankylosing spondylitis 90% of sufferers of which express the HLA-B27 antigen, versus 7% in the normal population

REAGENT COMPOSITION

The purified monoclonal HLA-B27 Antibody conjugated with fluorescein isothiocyanate (FITC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

• Clone: HLA–ABC–m3.
• Isotype: IgG2a
• Amount per 1 ml vial: 200 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody HLA-DR-FITC, Clone L243

• Antibody HLA-DR-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 HLA-DR antigenexpressing cells. This reagent must be used by flow cytometry qualified personal.

SUMMARY AND EXPLANATION

The main function of human leucocyte antigen (HLA) molecules is to present antigenic peptides to the T-cell receptor, thereby regulating the induction of the immune response. The HLA molecules are encoded by a cluster of tightly linked genes located on the short arm of chromosome 6. Three classes of HLA molecules (I, II and III) have been denoted. Human class II genes are located in the HLA-D region, consisting of three families called DQ, DP and DR. The products of class II genes form a heterodimeric transmembrane protein, consisting of a heavy α-chain and a light βchain. The DR α-chain is expressed from one non-polymorphic gene, whereas the DR β- chain originates from nine highly polymorphic genes. HLA-DR antigen is constitutively expressed on antigen-presenting cells, such as B lymphocytes, monocytes and dendritic cells but can also be detected on activated T lymphocytes and activated granulocytes. The antigen has been found expressed in cases of different types of acute lymphoblastic leukaemias, acute myeloid leukaemias, chronic lymphoblastic leukaemias, chronic myeloid leukaemias and B- and T-cell non-Hodgkin’s leukaemias. However, the antigen is normally not present on non-haematopoietic tumours and multiple myelomas.

REAGENT COMPOSITION

The purified monoclonal HLA-DR antibody conjugated with fluorescein isothiocyanate (FITC), is supplied in phosphatebuffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: L243
• Isotype: IgG2a.
• Amount per 1 mL vial: 200 tests (5 µL mAb per determination).
• Reagent is considered non-sterile

Name: Flow Cytometry Antibody HLA-DR-Pacific Blue^TM, Clone L243

• HLA-DR is a monoclonal antibody (mAb) labelled with Pacific Blue^TM designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of HLA-DR 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. The main function of human leucocyte antigen (HLA) molecules is to present antigenic peptides to the T-cell receptor, thereby regulating the induction of the immune response. The HLA molecules are encoded by a cluster of tightly linked genes located on the short arm of chromosome 6. Three classes of HLA molecules (I, II and III) have been denoted. Human class II genes are located in the HLA-D region, consisting of three families called DQ, DP and DR. The products of class II genes form a heterodimeric transmembrane protein, consisting of a heavy α-chain and a light βchain. The DR α-chain is expressed from one non-polymorphic gene, whereas the DR β- chain originates from nine highly polymorphic genes. HLA-DR antigen is constitutively expressed on antigen-presenting cells, such as B lymphocytes, monocytes and dendritic cells but can also be detected on activated T lymphocytes and activated granulocytes. The antigen has been found expressed in cases of different types of acute lymphoblastic leukaemias, acute myeloid leukaemias, chronic lymphoblastic leukaemias, chronic myeloid leukaemias and B- and T-cell non-Hodgkin’s leukaemias. However, the antigen is normally not present on non-haematopoietic tumours and multiple myelomas.

REAGENT COMPOSITION

The purified monoclonal HLA-DR Antibody conjugated with Pacific BlueTM is supplied in phosphate-buffered saline (PBS) containing 1% (m/v) BSA and 0.09% (m/v) sodium azide.

• Clone: L243
• Isotype: IgG2a.
• Amount per vial: 100 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody Kappa-FITC, Clone Polyclonal

• Kappa-FITC is a polyclonal antibody (pAb) labelled with fluorescein isothiocyanate (FITC) and designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells that express human Kappa immunoglobulin Light Chains by flow cytometry. Antibodies to Kappa Light Chains are useful for the identification of clonal excess in B-cell lymphoproliferative disorders together with a panel of other antibodies. Anti-Kappa Light Chains reacts with human free Kappa chains as well as Kappa chains in intact immunoglobulin molecules. This reagent must be used by flow cytometry qualified personal.       

SUMMARY AND EXPLANATION

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 cells (NK). B lymphocytes are the producers of antibodies and mediate humoral immunity particularly effective against toxins, whole bacteria, and free viruses. Most B cells, with the exception of pre-B progenitors, pre-B cells, and mature plasma cells, express immunoglobulin on their surface. Each cell expresses only one light chain type (Kappa or Lambda). In normal peripheral blood and lymph nodes, there is a mixture of Kappa positive and Lambda positive cells, with two-thirds of the cells expressing Kappa and one-third expressing Lambda. Since lymphoid neoplasms are usually clonal expansions of a single cell, malignant cells uniformly express the same Light Chain isotype. Neoplastic B cell lymphoprolipherative disorders can frequently be suspected based on the demonstration of a marker predominance of cells expressing a single Light Chain type. The Kappa immunoglobulin Light Chain count is generally expressed as a percentage of B cells present in the sample which can be determined by flow cytometry based on its positive expression of CD19. Since every flow cytometer has different operating characteristics each laboratory must determine its optimal operating procedure and the combination of markers used to identify the target cell population

REAGENT COMPOSITION

Purified polyclonal antibody anti-human Kappa Light Chains, Goat F(ab’)2, conjugated with fluorescein isothiocyanate (FITC), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide.

 Amount per 0,5 ml vial: 100 tests (5µl pAb to 106 cells).

Reagents are not considered sterile.