Product datasheet Anti-CD75 antibody [LN1] ab77676 3 Images Overview Product name Anti-CD75 antibody [LN1] Description Mouse monoclonal [LN1] to CD75 Specificity This antibody is shown to be a reliable antibody for ascribing a B-cell phenotype in known lymphoid tissues. We have data to indicate that this antibody may not cross react with Rat. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues. Tested applications IHC-P, Flow Cyt, ICC/IF Species reactivity Reacts with: Human Immunogen Tissue/ cell preparation - Nuclei from pokeweed mitogen-stimulated peripheral blood lymphocytes. Positive control Lymph node or human tonsil. Properties Form Liquid Storage instructions Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. Storage buffer Preservative: None Constituents: 10mM PBS, pH 7.4 Purity Ammonium Sulphate Precipitation Clonality Monoclonal Clone number LN1 Isotype IgM Light chain type kappa Applications Our Abpromise guarantee covers the use of ab77676 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. Application Abreviews Notes 1 Application Abreviews IHC-P Notes 1/25 - 1/50. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 min. Use 1µg for 106 cells. ab18400-Mouse monoclonal IgM, is suitable for use as an Flow Cyt isotype control with this antibody. ICC/IF Use at an assay dependent concentration. Target Function Transfers sialic acid from the donor of substrate CMP-sialic acid to galactose containing acceptor substrates. Pathway Protein modification; protein glycosylation. Sequence similarities Belongs to the glycosyltransferase 29 family. Post-translational modifications The soluble form derives from the membrane form by proteolytic processing. The HB-6, CDW75, and CD76 differentiation antigens are cell-surface carbohydrate determinants generated by this enzyme. Cellular localization Golgi apparatus > Golgi stack membrane. Secreted. Membrane-bound form in trans cisternae of Golgi. Secreted into the body fluid. Anti-CD75 antibody [LN1] images Formalin-fixed, paraffin-embedded human tonsil stained with ab77676 at 4-8ug/ml, using a peroxidase-conjugate secondary and AEC chromogen. Note cell membrane and cytoplasmic staining of B cells in germinal center. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CD75 antibody [LN1] - BSA and Azide free (ab77676) 2 ICC/IF image of ab77676 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBSTween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77676, 10µg/ml) overnight at +4°C. The secondary antibody Immunocytochemistry/ Immunofluorescence- (green) was ab96879, DyLight® 488 goat CD75 antibody [LN1] - BSA and Azide anti-mouse IgG (H+L) used at a 1/250 dilution free(ab77676) for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. Overlay histogram showing Ramos cells stained with ab77676 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBSTween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the Flow Cytometry-Anti-CD75 antibody [LN1] - BSA antibody (ab77676, 1µg/1x106 cells) for 30 and Azide free(ab77676) min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. 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