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 4 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 PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Left Panel) or Alexa Fluor® 647 Mouse Anti-XBP-1S antibody (Cat. No. 562821). 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.
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 4 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 PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Left Panel) or Alexa Fluor® 647 Mouse Anti-XBP-1S antibody (Cat. No. 562821). 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.
製品詳細
BD Pharmingen™
XBP1; X-box binding protein 1; TREB5; Tax-responsive element-binding
Mouse (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Human XBP-1S Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2737817
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 monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
関連製品
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ
0.1 mg
カタログ番号
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ
0.5 mg
カタログ番号
553142
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Transcription Factor Buffer Set RUO
サイズ
100 Tests
カタログ番号
562574
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
サイズ
100 Tests
カタログ番号
557732
Transcription Factor Buffer Set RUO
サイズ
25 Tests
カタログ番号
562725
562821 Rev. 1
抗体の詳細
Q3-695
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.
562821 Rev. 1
フォーマットの詳細
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 520-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
562821 Rev.1
引用＆参考文献
Development References (3)
-
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
-
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
-
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
562821 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products 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.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
