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Alexa Fluor™ 647 Mouse Anti-Human SCIMP
Alexa Fluor™ 647 Mouse Anti-Human SCIMP
Two-color flow cytometric analysis of SCIMP expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stained with FITC Mouse Anti-Human HLA-DR, DP, DQ (Cat. No. 555558/550853/562008) and then washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656]. After the cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No 554723), they were stained with either Alexa Fluor™647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365, Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human SCIMP antibody (Cat. No. 568780/568955; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of SCIMP (or Ig Isotype control staining) versus HLA-DR, DP, DQ was derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of SCIMP expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stained with FITC Mouse Anti-Human HLA-DR, DP, DQ (Cat. No. 555558/550853/562008) and then washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656]. After the cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No 554723), they were stained with either Alexa Fluor™647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365, Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human SCIMP antibody (Cat. No. 568780/568955; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of SCIMP (or Ig Isotype control staining) versus HLA-DR, DP, DQ was derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
SLP adaptor and CSK interacting membrane protein
Human (QC Testing)
Mouse IgG2a, κ
Human Recombinant intracellular part of human SCIMP.
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
388325
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO


Preparation And Storage

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 and unreacted dye was removed.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. For U.S. patents that may apply, see bd.com/patents.
568780 Rev. 2
Antibody Details
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NVL-07

The NVL-07 monoclonal antibody specifically recognizes SLP adaptor and CSK-Interacting Membrane Protein (SCIMP), also known as SLP adapter.  SCIMP is a lipid tetraspanin-associated transmembrane adaptor protein that belongs to the palymitoylated transmembrane adaptor protein (pTRAP) family. SCIMP localizes to the immunological synapse and contains a short extracellular domain, a long intracellular domain with multiple tyrosine phosphorylation sites, a proline-rich motif, and two palmitoylation sites. SCIMP is expressed in antigen presenting cells, including B cells, dendritic cells, and macrophages of the lymph nodes and spleen.  This TLR signaling adaptor regulates antigen presentation; it is involved in (MHC) class II signaling transduction in B cells and Dectin-1 signaling in dendritic cells by acting as a membrane-bound scaffold for Src-family kinases. SCIMP participation in TLR signaling yields proinflammatory cytokines Il-6 and Il-12p40 in macrophages, and the inflammasome-driven Il-1β. As an important regulator of host immune responses, SCIMP expression levels are studied in autoimmune and inflammatory diseases.

568780 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
568780 Rev.2
Citations & References
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View product citations for antibody "568780" on CiteAb

Development References (6)

  1. Curson JEB, Luo L, Sweet MJ, Stow JL. pTRAPs: Transmembrane adaptors in innate immune signaling.. J Leukoc Biol. 2018. (Biology). View Reference
  2. Draber P, Vonkova I, Stepanek O, et al. SCIMP, a transmembrane adaptor protein involved in major histocompatibility complex class II signaling.. Mol Cell Biol. 2011; 31(22):4550-62. (Immunogen: Immunoprecipitation, Western blot). View Reference
  3. Lucas RM, Liu L, Curson JEB, et al. SCIMP is a spatiotemporal transmembrane scaffold for Erk1/2 to direct pro-inflammatory signaling in TLR-activated macrophages.. Cell Rep. 2021; 36(10):109662. (Biology). View Reference
  4. Luo L, Bokil NJ, Wall AA, et al. SCIMP is a transmembrane non-TIR TLR adaptor that promotes proinflammatory cytokine production from macrophages.. Nat Commun. 2017; 8:14133. (Biology). View Reference
  5. Luo L, Curson JEB, Liu L, et al. SCIMP is a universal Toll-like receptor adaptor in macrophages.. J Leukoc Biol. 2020; 107(2):251-262. (Biology). View Reference
  6. Luo L, Lucas RM, Liu L, Stow JL. Signalling, sorting and scaffolding adaptors for Toll-like receptors.. J Cell Sci. 2019; 133(5):jcs239194. (Biology). View Reference
View All (6) View Less
568780 Rev. 2

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.