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Two-color flow cytometric analysis of XBP-1S expression in activated mouse splenocytes. Mouse splenic leucocytes were stimulated with lipopolysaccharide (LPS; Sigma L-3137; 1 μg/ml) for 3 days (37°C). Cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon™ BV421 Mouse Anti-XBP-1S antibody (Cat. No. 563382). Two-color flow cytometric dot plots show the correlated expression of CD45R/B220 versus XBP-1S (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of intact activated splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-XBP-1S
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The Q3-695 monoclonal antibody specifically binds to XBP-1S, the spliced isoform of X-box binding protein 1 (XBP-1). XBP-1S is generated by cells that undergo an unfolded protein response, ie, cells responding to the accumulation of unfolded proteins in the endoplasmic reticulum. It plays important roles as a transcription factor in a number of important cellular processes including the regulation of MHC class II genes, differentiation of plasma cells, immunoglobulin secretion and hepatocyte growth.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.
Development References (3)
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Gass JN, Gifford NM, Brewer JW. Activation of an unfolded protein response during differentiation of antibody-secreting B cells. J Biol Chem. 2002; 277(50):49047-49054. (Biology). View Reference
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Iwakoshi NN, Lee AH, Vallabhajosyula P, Otipoby KL, Rajewsky K, Glimcher LH. Plasma cell differentiation and the unfolded protein response intersect at the transcription factor XBP-1. Nat Immunol. 2003; 4(4):321-329. (Biology). View Reference
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Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K. XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell. 2001; 107(7):881-891. (Biology). 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.
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