Description
The 1A5 monoclonal antibody specifically recognizes Sialic acid binding Ig-like lectin 5 (Siglec-5) which is also known as CD170, CD33 antigen-like 2 (CD33L2) and Obesity binding protein-2 (OBBP2). CD170 is a type I transmembrane glycoprotein that forms a ~140 kDa dimer on the cell surface. CD170 is encoded by SIGLEC5 which belongs to the SIGLEC (sialic acid binding Ig-like lectin) family within the immunoglobulin superfamily. The extracellular region of CD170 consists of an N-terminal V-set Ig-like domain, which contains the sialic acid binding site, followed by 3 C2-set Ig like domains, a transmembrane region and a cytoplasmic tail containing 2 immunoreceptor tyrosine-based inhibitory motifs (ITIMs) which regulate cellular signaling. CD170 bind to alpha-2,3- and alpha-2,6-linked sialic acid glycoproteins and glycolipids and to glycophorin A. It is highly expressed on neutrophils and expressed at lower levels on monocytes, macrophages, dendritic cells, and some lymphocytes. CD170 is an adhesion molecule that may function in regulating cellular activation and in sialic-acid dependent binding to cells.
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.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. 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.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. 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.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. 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.
11. 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.
12. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
13. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Specifications

General

BrandBD OptiBuild™
Alternative NameSIGLEC-5; SIGLEC5; CD33 antigen-like 2; CD33L2; obesity-binding protein 2; OBBP2
ReactivityHuman (Tested in Development)
IsotypeMouse BALB/c IgG1, κ
ImmunogenHuman Siglec-5 Recombinant Protein
ApplicationFlow cytometry (Qualified)
Concentration0.2 mg/ml
Workshop NumberVII 70443
Entrez Gene ID8778
RRIDAB_3693592
Storage BufferAqueous buffered solution containing ≤0.09% sodium azide.
Regulatory StatusRUO