The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 2µg for 106 cells.
ab35768 - Rat monoclonal IgM, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. Dilutions have to be made according to the amounts of SIGN-R1 to be inactivated.
SIGN R1 is a specific marker for the identification of macrophage subpopulations present in the marginal zone of spleen (the so-called marginal zone macrophages (MZM)), in the lymph node medulla, and in some strains, in the peritoneal cavity. MZM of the spleen are involved in the clearance of polysaccharides.
Mouse SIGN R1 is a C type lectin, like DC SIGN which is expressed on Human dendritic cells (DCs). However, Mmouse SIGN R1 itself is not expressed on DCs. SIGN R1 exists in an aggregated form, resistant to dissociation into monomers upon boiling in SDS under reducing conditions.
SIGN R1 mediates the uptake of encapsulated organisms and may be an important mediator for the uptake of microbes in both spleen and lymph node, particularly through the recognition of microbial polysaccharides.
Immunohistochemistry (Frozen sections) - Anti-SIGN Related 1 antibody [ER-TR9] (ab37220)This image is courtesy of an Abreview by Lynda Coughlan.
ab37220 staining SIGN Related 1 in Mouse spleen tissue sections by Immunohistochemistry (Frozen sections). The sections were fixed with 4% PFA for 10 minutes at room temperature prior to blocking with 10% Goat serum for 30 minutes at 15°C. The primary antibody was diluted to a concentration of 4 µg/mL in PBS with 2% Goat serum and incubated with the sample for 1 hour at 15°C. An Alexa Fluor® 488-conjugated Goat anti-Rat IgM polyclonal was used as the secondary antibody, diluted 1/500.
Flow Cytometry - Anti-SIGN Related 1 antibody [ER-TR9] (ab37220)
Overlay histogram showing SP0/2 cells stained with ab37220 (red line). The cells were fixed with 4% paraformaldehyde (10min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37220, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgM, kappa monoclonal [RTK2118] (ab35768, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SP0/2 cells fixed with 80% methanol (5 min) used under the same conditions. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
Taylor PR et al. The role of SIGNR1 and the beta-glucan receptor (dectin-1) in the nonopsonic recognition of yeast by specific macrophages. J Immunol172:1157-62 (2004).
Blocking, Flow Cyt
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Kretschmer K et al. The selection of marginal zone B cells differs from that of B-1a cells. J Immunol171:6495-501 (2003).
Read more (PubMed: 14662849) »