BD Horizon™ BV421 Mouse Anti-Human VSTM1 (SIRL-1)
Clone 1A5/SIRL-1 (also known as 1A5) (RUO)
Multiparameter flow cytometric analysis of VSTM1 (SIRL-1) expression on different Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human VSTM1 (SIRL-1) antibody (Cat. No. 569948/569954; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of VSTM1 (SIRL-1) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of VSTM1 (SIRL-1) expression on different Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human VSTM1 (SIRL-1) antibody (Cat. No. 569948/569954; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of VSTM1 (SIRL-1) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Preparation And Storage
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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The 1A5 (1A5/SIRL-1) monoclonal antibody specifically recognizes human V-set and transmembrane domain-containing protein 1 (VSTM1) which is also known as Signal Inhibitory Receptor on Leucocytes-1 (SIRL-1). VSTM1 (SIRL-1) is a single-pass type I transmembrane glycoprotein that contains an IgV-like domain in its extracellular region which is followed by a transmembrane segment and an intracellular region with two immunoreceptor tyrosine-based inhibitory motifs (ITIMs). This molecule is encoded by VSTM1 (V-set and transmembrane domain containing 1) which belongs to the transmembrane receptor Ig superfamily of immune inhibitory receptors. VSTM1 (SIRL-1) is variably expressed on myeloid cells including neutrophils, eosinophils, and monocytes. SIRL-1 recognizes phenol-soluble modulins of Staphylococcus as well as danger- or damage-associated molecular patterns (DAMPs) including S100 proteins and the antimicrobial peptide, LL-37. VSTM1 (SIRL-1) regulates innate effector functions including Fc Receptor-induced production of reactive oxygen species (ROS) and formation of neutrophil extracellular traps (NET) released by human neutrophils in response to certain bacterial and inflammatory danger signals. Although NETs serve to trap and kill extracellular microbes, VSTM1 (SIRL-1)-mediated regulation of NETs expression helps avoid excessive tissue damage caused by inflammation and potential induction or exacerbation of autoimmune disease. VSTM1 (SIRL-1) function does not impair neutrophil phagocytosis and killing of intracellular bacteria. VSTM1 (SIRL-1) expression is downregulated in response to microbial stimuli which allows microbial killing and clearance of pathogens.
Development References (4)
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Rumpret M, von Richthofen HJ, Peperzak V, Meyaard L. Inhibitory pattern recognition receptors.. J Exp Med. 2022; 219(1):e20211463. (Biology). View Reference
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Steevels TA, Lebbink RJ, Westerlaken GH, Coffer PJ, Meyaard L. Signal inhibitory receptor on leukocytes-1 is a novel functional inhibitory immune receptor expressed on human phagocytes.. J Immunol. 2010; 184(9):4741-8. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation, Western blot). View Reference
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Steevels TA, van Avondt K, Westerlaken GH, et al. Signal inhibitory receptor on leukocytes-1 (SIRL-1) negatively regulates the oxidative burst in human phagocytes.. Eur J Immunol. 2013; 43(5):1297-308. (Clone-specific: Flow cytometry). View Reference
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Van Avondt K, van der Linden M, Naccache PH, Egan DA, Meyaard L. Signal Inhibitory Receptor on Leukocytes-1 Limits the Formation of Neutrophil Extracellular Traps, but Preserves Intracellular Bacterial Killing.. J Immunol. 2016; 196(9):3686-94. (Clone-specific: Functional assay). View Reference
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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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
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.