Flow cytometric analysis of SSEA-5 expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 48 grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527) and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human SSEA-5 monoclonal antibody (solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of the H9 human ES cell line. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.
Immunoflourescent staining of SSEA-5 on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 43 grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC- qualified Matrix (Cat. No. 354277) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). Cells were stained with Alexa Fluor® 647 Mouse Anti-Human SSEA-5 monoclonal antibody (pseudo-colored red) at 2.5 μg/mL. Cell nuclei were stained with DAPI (pseudo- colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.
Flow cytometric analysis of SSEA-5 expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 48 grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527) and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human SSEA-5 monoclonal antibody (solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of the H9 human ES cell line. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.
Immunoflourescent staining of SSEA-5 on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 43 grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC- qualified Matrix (Cat. No. 354277) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). Cells were stained with Alexa Fluor® 647 Mouse Anti-Human SSEA-5 monoclonal antibody (pseudo-colored red) at 2.5 μg/mL. Cell nuclei were stained with DAPI (pseudo- colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.
製品詳細
BD Pharmingen™
Human (QC Testing)
Mouse IgG1, κ
Human Undifferentiated Embryonic Stem Cells
Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_2738404
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 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.
- 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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- mTESR™1 is a trademark of StemCell Technologies.
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- Patent Pending.
- 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
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Fixation Buffer RUO
サイズ
100 mL
カタログ番号
554655
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
サイズ
100 Tests
カタログ番号
557732
Accutase™ Cell Detachment Solution RUO
サイズ
100 mL
カタログ番号
561527
563753 Rev. 1
抗体の詳細
8e11/SSEA-5
Stage-specific embryonic antigen (SSEA)-5 is a pluripotency surface marker expressed in the blastocyst inner cell mass and on human pluripotent stem cells, including both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Because SSEA-5 expression rapidly decreases upon differentiation, it can be used to identify undifferentiated pluripotent stem cells. SSEA-5 can be combined with other pluripotency surface markers (e.g. CD9/CD90 or CD50/CD200) to immunodeplete remaining pluripotent stem cells from incompletely differentiated cultures.
563753 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
563753 Rev.1
引用＆参考文献
Development References (1)
-
Tang C, Lee AS, Volkmer JP, et al. An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells. Nat Biotechnol. 2011. (Clone-specific: Blocking, Depletion, Immunofluorescence). View Reference
563753 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.
