BD Horizon™ RB780 Rat Anti-Mouse CD169 (Siglec-1)
Clone 3D6/CD169 (also known as 3D6) (RUO)
Multicolor flow cytometric analysis of CD169 (Siglec -1) expression on Mouse myeloid cells. C57BL/6 Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 553128) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse CD169 (Siglec-1) antibody (Cat. No. 570772/570850; Right Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD169 (Siglec-1) [or Ig Isotype control staining] versus Ly-6G and Ly-6C was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
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Multicolor flow cytometric analysis of CD169 (Siglec -1) expression on Mouse myeloid cells. C57BL/6 Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 553128) and with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; Left Plot) or BD Horizon™ RB780 Rat Anti-Mouse CD169 (Siglec-1) antibody (Cat. No. 570772/570850; Right Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD169 (Siglec-1) [or Ig Isotype control staining] versus Ly-6G and Ly-6C was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
Anti-mouse sialoadhesin
Mouse (QC Testing)
Rat AO IgG2a, κ
Purified sialoadhesin from mouse spleen
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
RB780 Rat IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
568698
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DAPI Solution RUO
Size
1 mg
Cat No.
564907
PE Rat Anti-Mouse Ly-6G and Ly-6C RUO
Size
0.1 mg
Cat No.
553128
570850 Rev. 1
Antibody Details
3D6/CD169
The 3D6 monoclonal antibody specifically recognizes CD169 which is a ~185 kDa type I transmembrane glycoprotein that is encoded by Siglec1. CD169 belongs to the Sialic acid binding Ig like lectins (Siglec) family and is also known as Sialoadhesin (Sn), Siglec-1, or Sheep erythrocyte receptor (SER). CD169 is expressed on subsets of macrophages found in the bone marrow, secondary lymphoid tissues, liver colon and lungs. It binds to the oligosaccharide sequence NeuAc alpha 2,3Gal present on glycolipids and glycoproteins including CD43, CD162, CD206, or CD227. CD169 serves as an adhesion molecule that can mediate cellular interactions with the extracellular matrix or with other cells including lymphocytes and myeloid cells. CD169 may play a role in erythroid and myeloid cell development as well as leucocyte trafficking and inflammation.
570850 Rev. 1
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
570850 Rev.1
Citations & References
View product citations for antibody "570850" on CiteAb
Development References (4)
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Crocker PR, Freeman S, Gordon S, Kelm S. Sialoadhesin binds preferentially to cells of the granulocytic lineage.. J Clin Invest. 1995; 95(2):635-43. (Immunogen: Blocking, Radioimmunoassay). View Reference
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Ducreux J, Crocker PR, Vanbever R. Analysis of sialoadhesin expression on mouse alveolar macrophages.. Immunol Lett. 2009; 124(2):77-80. (Clone-specific: Blocking, Inhibition). View Reference
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Kumamoto Y, Higashi N, Denda-Nagai K, et al. Identification of sialoadhesin as a dominant lymph node counter-receptor for mouse macrophage galactose-type C-type lectin 1.. J Biol Chem. 2004; 279(47):49274-80. (Clone-specific: Fluorescence microscopy, Immunofluorescence, Western blot). View Reference
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Nath D, van der Merwe PA, Kelm S, Bradfield P, Crocker PR. The amino-terminal immunoglobulin-like domain of sialoadhesin contains the sialic acid binding site. Comparison with CD22.. J Biol Chem. 1995; 270(44):26184-91. (Clone-specific: Functional assay, Immunoaffinity chromatography). View Reference
570850 Rev. 1
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.