CD170 (Siglec F) Monoclonal Antibody (1RNM44N), Brilliant Violet™ 480, eBioscience™

Product Specific Information

Description
This 1RNM44N monoclonal antibody recognizes mouse CD170, also known as Siglec F. Mouse Siglec F is a functional ortholog of human Siglec 8, however, unlike human Siglec 8, mouse Siglec F is not expressed on mast cells. This 1RNM44N antibody will recognize a formaldehyde-fixed epitope.

Applications Tested
This 1RNM44N antibody has been tested by flow cytometric analysis of mouse thioglycolate-elicited peritoneal exudate cells. This may be used at less than or equal to 1.0 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
• Our internal testing suggests that Brilliant Violet™ 480 (BV480) is not compatible with methanol-based fixation.

Excitation: 440 nm; Emission: 479 nm; Laser: Violet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

Target Information

SIGLEC5 is a putative adhesion molecule that mediates sialic-acid dependent binding to cells. This protein binds equally to alpha2,3-linked and alpha2,6-linked sialic acid. The sialic acid recognition site may be masked by cis interactions with sialic acids on the same cell surface. This molecule is expressed by monocytic/myeloid lineage cells. SIGLEC5 is found at high levels in peripheral blood leukocytes, spleen, bone marrow and at lower levels in lymph node, lung, appendix, placenta, pancreas and thymus. It is expressed by monocytes and neutrophils but absent from leukemic cell lines representing early stages of myelomonocytic differentiation.