فلوسایتومتری

Name: Flow Cytometry Antibody CD138-PECy5, Clone B-A38

• Antibody CD138-PECy5 is a monoclonal antibody (mAb) labelled with R-phycoerythrin-cyanine 5 (PECy5) designed for flow cytometry (FC) use as a direct immunofluorescence reagent in the identification and enumeration of CD138 antigenexpressing cells.        

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. The CYT-138C4 mAb reacts with 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.

REAGENT COMPOSITION

The purified monoclonal CD138 Antibody conjugated with R-phycoerythrin-cyanine 5 (PECy5) is supplied in phosphatebuffered saline (PBS) containing 0,09% sodium azide.

• Clone: B-A38.
• Isotype: IgG1.
• Amount per 1 mL vial: 200 tests (5 µL mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody CD180-APC-C750, Clone G28-8

• Antibody CYT-180AC750 is a monoclonal antibody (mAb) labelled with tandem allophycocyanin-C750 (APC-C750^TM) designed for use as a direct immunofluorescence reagent in the identification and enumeration of cells which express the CD180 antigen by flow cytometry. This reagent must be used by flow cytometry qualified personnel.

SUMMARY AND EXPLANATION

CD180 (RP105), belongs to the toll-like receptor (TLR) family and is expressed on most B cells, monocytes/macrophages, and dendritic cells. It is a 105 kDa protein consisting of extracellular leucine-rich repeats and a short cytoplasmic tail. MD-1, a secretory protein, is associated with CD180 at the cell surface and, in conjunction with TLR4, recognizes bacterial lipopolysaccharide. Ligation of CD180 by monoclonal antibodies leads to B cell activation.

REAGENT COMPOSITION

Purified monoclonal CD180 antibody conjugated with tandem allophycocyanin-C750 (APC-C750TM), supplied in phosphate buffered saline with 0,09% sodium azide.

• Clone: G28-8.
• Isotype: Mouse / IgG1.
• Amount per vial: 50 tests. 3 µl mAb per determination.
• Reagents are not considered sterile.

Name: Flow Cytometry Antibody CD185-APC, Clone MU5UBEE

• Antibody CD185-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 CD185 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. CD185, also known as C-X-C chemokine receptor 5 (CXCR5) and Burkitt lymphoma receptor 1 (BLR1), is a 7 transmembrane domain protein mainly expressed on B cells and CD4+ T cells (1, 2). CD185 has been selected by EuroFlow consortium in its B-CLPD (B-cell chronic lymphoproliferative diseases) panel.

REAGENT COMPOSITION

The purified monoclonal CD185 antibody conjugated with allophycocyanin (APC) is supplied in phosphate-buffered saline (PBS) containing 0.1% sodium azide.

• Clone: MU5UBEE.
• Isotype: IgG2b.
• Amount per vial: 100 tests (5 µl mAb per determination).
• Reagent is considered non-sterile.

Name: Flow Cytometry Antibody IgA1-APC, Clone SAA1

IgA1-APC is designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of IgA1 expressing cells. IgA1-APC reacts with the Fc portion of the heavy chain of human IgA1.

In humans, 85–90% of IgA antibodies are found in body secretions such us saliva, tears, breast milk; gastrointestinal mucous membranes, urogenital and respiratory tracts and prostatic fluid, among others. The remaining fraction of 10–15% of IgA antibodies is found in blood, produced by B cells in the lymph nodes, bone marrow, and spleen. Two subclasses of human serum IgA exist. IgA1 tends to be monomeric and is present at a fivefold higher concentration than IgA2. Soluble IgA2 tends to be present more often in polymeric forms.

CYT-IGA1AP should be used with BulkLysisTM (CYT-BL) buffer that allows you to stain a great number of cells and analyze low frequency subsets, as IgA1+ memory B cells and plasma cells. If another lysing protocol is used, two pre-staining washing steps using PBS + 1% (m/v) BSA + 0,09% (m/v) sodium azide must be performed. This reagent must be used by flow cytometry qualified personnel.

REAGENT COMPOSITION

Purified monoclonal anti-IgA1 antibody (mAb) conjugated with allophycocyanine (APC), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide and BSA 1% (m/v).

Clone: SAA1.

Isotype: Mouse IgG1.

Amount per vial: 25 tests using 3µL of mAb to 106 cells.

Name: Flow Cytometry Antibody IgA1-PE, Clone SAA1

• IgA1-PE is designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of IgA1 expressing cells. IgA1-PE reacts with the Fc portion of the heavy chain of human IgA1.

In humans, 85–90% of IgA antibodies are found in body secretions such us saliva, tears, breast milk, gastrointestinal mucous membranes, urogenital and respiratory tracts and prostatic fluid, among others. The remaining fraction of 10–15% of IgA antibodies is found in blood, produced by B cells in the lymph nodes, bone marrow, and spleen. Two subclasses of human serum IgA exist. IgA1 tends to be monomeric and is present at a fivefold higher concentration than IgA2. Soluble IgA2 tends to be present more often in polymeric forms.

 CYT-IGA1PE should be used with BulkLysis^TM (CYT-BL) buffer that allows you to stain a great number of cells and analyze low frequency subsets, as IgA1+ memory B cells and plasma cells. If another lysing protocol is used, two pre-staining washing steps using PBS + 1% (m/v) BSA + 0,09% (m/v) sodium azide must be performed. This reagent must be used by flow cytometry qualified personnel.

REAGENT COMPOSITION

Purified monoclonal anti-IgA1 antibody (mAb) conjugated with R-phycoerythrin (PE), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide and BSA 1% (m/v).

Clone: SAA1.

Isotype: Mouse IgG1.

Amount per vial: 25 tests using 3µL of mAb to 106 cells.

Name: Flow Cytometry Antibody IgA1-PerCP-Cyanine5.5, Clone SAA1

Antibody IgA1-PerCP-Cyanine5.5 is designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of IgA1 expressing cells. IgA1-PerCP-Cyanine5.5 reacts with the Fc portion of the heavy chain of human IgA2.

In humans, 85–90% of IgA antibodies are found in body secretions such us saliva, tears, breast milk; gastrointestinal mucous membranes, urogenital and respiratory tracts and prostatic fluid, among others. The remaining fraction of 10–15% of IgA antibodies is found in blood, produced by B cells in the lymph nodes, bone marrow, and spleen. Two subclasses of human serum IgA exist. IgA1 tends to be monomeric and is present at a fivefold higher concentration than IgA2. Soluble IgA2 tends to be present more often in polymeric forms.

CYT-IGA1C should be used with BulkLysis^TM (CYT-BL) buffer that allows you to stain a great number of cells and analyze low frequency subsets, as IgA1+ memory B cells and plasma cells. If another lysing protocol is used, two pre-staining washing steps using PBS + 1% (m/v) BSA + 0,09% (m/v) sodium azide must be performed.

REAGENT COMPOSITION

Purified monoclonal anti-IgA1 antibody (mAb) conjugated with Peridin chlorophyll protein-Cyanine 5.5 (PerCP-Cyanine5.5), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide and BSA 1% (m/v).

• Clone: SAA1.
• Isotype: Mouse IgG1.
• Amount per vial: 25 tests using 3µL of mAb to 106 cells.

Name: Flow Cytometry Antibody IgA2-PerCP-Cyanine5.5, Clone SAA2

Antibody IgA2-PerCP-Cyanine5.5 is designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of IgA2 expressing cells. IgA2-PerCP-Cyanine5.5 reacts with the Fc portion of the heavy chain of human IgA2.

In humans, 85–90% of IgA antibodies are found in body secretions such us saliva, tears, breast milk; gastrointestinal mucous membranes, urogenital and respiratory tracts and prostatic fluid, among others. The remaining fraction of 10–15% of IgA antibodies is found in blood, produced by B cells in the lymph nodes, bone marrow, and spleen. Two subclasses of human serum IgA exist. IgA1 tends to be monomeric and is present at a fivefold higher concentration than IgA2. Soluble IgA2 tends to be present more often in polymeric forms.

CYT-IGA2C2 should be used with BulkLysis^TM (CYT-BL) buffer that allows you to stain a great number of cells and analyze low frequency subsets, as IgA2+ memory B cells and plasma cells. If another lysing protocol is used, two pre-staining washing steps using PBS+1% (m/v) BSA + 0,09% (m/v) sodium azide must be performed.

REAGENT COMPOSITION

Purified monoclonal anti-IgA2 antibody (mAb) conjugated with Peridin chlorophyll protein-Cyanine 5.5 (PerCP-Cyanine5.5), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide and BSA 1% (m/v).

• Clone: SAA2.
• Isotype: Mouse IgG1.
• Amount per vial: 25 tests using 3µL of mAb to 106 cells

Name: Flow Cytometry Antibody IgD-PerCP-Cyanine5.5  , Clone IADB6.

• Antibody IgD-PerCP-Cyanine5.5 is designed for flow cytometry use as a direct immunofluorescence reagent in the identification and enumeration of IgD expressing cells. IgD-PerCP-Cyanine5.5 reacts with the delta chain of human IgD. IgD is barely detectable in the blood (0,001% of total serum Ig) and is rarely secreted. It is observed mostly on the surface of immature, peripheral B cells that already express IgM. The expression of IgD is transient; upon activation of the B cell by the binding of antigen, the surface expression of mIgD, but not mIgM, is abrogated.

CYT-IGDC should be used with BulkLysisTM (CYT-BL) buffer that allows you to stain a great number of cells and analyze low-frequency subsets, as IgD+ memory B cells and plasma cells. If another lysing protocol is used, two pre-staining washing steps using PBS + 1% (m/v) BSA + 0,09% (m/v) sodium azide must be performed.

REAGENT COMPOSITION

Purified monoclonal anti-IgD antibody (mAb) conjugated with Peridin chlorophyll protein-Cyanine 5.5 (PerCP-Cyanine5.5), supplied in phosphate buffered saline with 0,09% (m/v) sodium azide and BSA 1% (m/v).

• Clone: IADB6.
• Isotype: Mouse IgG2a.
• Amount per vial: 25 tests using 3µL of mAb to 106 cells

Name: Flow Cytometry Antibody CD81-APC-C750™ (RUO) Clone M38

• Anti-human CD81-APC-C750^TM is a monoclonal antibody (mAb) labelled with the tandem allophycocyanine-C750 (APCC750^TM) 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 personnel.

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 the tandem allophycocyanine-C750 (APC-C750TM), 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 0,15 mL vial: 50 tests (3 µL mAb per determination)
• Reagents are not considered sterile.