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BV480 Mouse Anti-Human IgM
BV480 Mouse Anti-Human IgM
Two-color flow cytometric analysis of IgM expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were incubated in complete medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Mouse IgG1, κ Isotype Control (Cat. No.565652; Left Plot) or BD Horizon™ BV480 Mouse Anti-Human IgM antibody (Cat. No. 566146/566153; Right Plot). Two-color flow cytometric dot plots showing the correlated expression of cell surface IgM (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of IgM expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were incubated in complete medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Mouse IgG1, κ Isotype Control (Cat. No.565652; Left Plot) or BD Horizon™ BV480 Mouse Anti-Human IgM antibody (Cat. No. 566146/566153; Right Plot). Two-color flow cytometric dot plots showing the correlated expression of cell surface IgM (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
IGHM; MU; Ig mu chain C region; AGM1; VH
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
3507
AB_2739544
Aqueous buffered solution containing BSA 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 antibody was conjugated with BD Horizon BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 were removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566146 Rev. 1
Antibody Details
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G20-127

IgM is an important component in the first line of defense against foreign pathogens, but may also play a role in autoimmune diseases.  IgM monomers consist of two light and two heavy chains.  Unlike the heavy chain of an IgG antibody which contains 3 constant Ig domains, the µ heavy chain of IgM contains 4 constant Ig domains.  Five IgM monomers complex with a small polypeptide (J-chain) to form pentameric IgM that can be found in human plasma.  In an immune response, the binding of IgM to a cell surface antigen enables C1q to activate interactions with downstream components in the classical complement pathway.  Mature B lymphocytes express IgM.  The G20-127 monoclonal antibody binds to the heavy chain of human IgM. The G20-127 antibody is not thought to react with other immunoglobulin heavy chain isotypes.

The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

566146 Rev. 1
Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV480
Violet 405 nm
440 nm
479 nm
566146 Rev.1
Citations & References
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View product citations for antibody "566146" on CiteAb

Development References (5)

  1. Chtanova T, Tangye SG, Newton R, et al. T follicular helper cells express a distinctive transcriptional profile, reflecting their role as non-Th1/Th2 effector cells that provide help for B cells. J Immunol. 2004; 173(1):68-78. (Clone-specific: Flow cytometry). View Reference
  2. Le Gallou S, Caron G, Delaloy C, Rossille D, Tarte K, Fest T. IL-2 requirement for human plasma cell generation: coupling differentiation and proliferation by enhancing MAPK-ERK signaling. J Immunol. 2012; 189(1):161-173. (Clone-specific: Flow cytometry). View Reference
  3. Mei HE, Yoshida T, Sime W, et al. Blood-borne human plasma cells in steady state are derived from mucosal immune responses. Blood. 2009; 113(11):2461-2469. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  4. Widhopf GF, 2nd, Brinson DC, Kipps TJ, Tighe H. Transgenic expression of a human polyreactive Ig expressed in chronic lymphocytic leukemia generates memory-type B cells that respond to nonspecific immune activation. J Immunol. 2004; 172(4):2092-2099. (Clone-specific: Flow cytometry). View Reference
  5. Zola H, Macardle PJ, Flego L, Webster J. The expression of sub-population markers on B cells: a re-evaluation using high-sensitivity fluorescence flow cytometry. Dis Markers. 1991; 9(2):103-118. (Biology: Cell differentiation, Dot Blot, Fluorescence activated cell sorting, In situ hybridization). View Reference
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566146 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.