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PE Mouse anti-Human TRA-1-60 Antigen
PE Mouse anti-Human TRA-1-60 Antigen
Flow cytometric analysis of PE Mouse Anti-Human TRA-1-60 on H9 cells.  H9 human embryonic stem (ES) cells were harvested with Accutase™ and stained with either PE Mouse Anti-Human TRA-1-60 (solid line) or PE mouse IgM (G155-228) isotype control (Cat. No. 555584, dashed line), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of PE Mouse Anti-Human TRA-1-60 on H9 cells.  H9 human embryonic stem (ES) cells were harvested with Accutase™ and stained with either PE Mouse Anti-Human TRA-1-60 (solid line) or PE mouse IgM (G155-228) isotype control (Cat. No. 555584, dashed line), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Flow cytometry was performed on a BD FACSCalibur™ System.
Product Details
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BD Pharmingen™
TRA-1-60(R)
Human (QC Testing), Rhesus (Reported)
Mouse BALB/c IgM, κ
Human Embryonal Carcinoma Cell Line
Flow cytometry (Routinely Tested)
20 µl
AB_1645539
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

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. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560193 Rev. 4
Antibody Details
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TRA-1-60

The TRA-1-60 monoclonal antibody reacts with the neuraminidase-resistant form of a pluripotent-stem-cell-specific epitope on a high-molecular-weight transmembrane glycoprotein.  The TRA-1-60 antigen is a sialylated epitope on the same keratan sulfate core molecule, podocalyxin, as 4 other distinct antigens on tumor-derived cell lines, TRA-1-81, GCTM2, K4, and K21.  The expression of TRA-1-60 antigen is stage-specific and can be used to characterize embryonic cells and monitor their differentiation.  The antigen is found on teratocarcinoma (embryonal carcinoma or EC), embryonic inner cell mass (but not morula or trophoblast), and embryonic stem (ES) cells.  TRA-1-60 antigen is released into the serum of patients bearing  testicular tumors containing EC cells.  As human EC and ES cells undergo differentiation, expression of TRA-1-60 antigen is lost.  Expression of TRA-1-60 antigen has also been observed on a rhesus monkey ES cell line (Thomson et al, 1995).

560193 Rev. 4
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
560193 Rev.4
Citations & References
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Development References (7)

  1. Andrews PW, Banting G, Damanov I, Arnaud D, Avner P. Three monoclonal antibodies defining distinct differentiation antigens associated with different high molecular weight polypeptides on the surface of human embryonal carcinoma cells. Hybridoma. 1984; 3(4):347-361. (Immunogen). View Reference
  2. Badcock G, Pigott C, Goepel J, Andrews PW. The human embryonal carcinoma marker antigen TRA-1-60 is a sialylated keratan sulfate proteoglycan. Cancer Res. 1999; 59:4715-4719. (Clone-specific: Immunoprecipitation). View Reference
  3. Draper JS, Pigott C, Thomson JA, Andrews PW. Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J Anat. 2002; 200:249-258. (Clone-specific: Flow cytometry). View Reference
  4. Henderson JK, Draper JS, Baillie HS, et al. Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens. Stem Cells. 2002; 20:329-337. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  5. Schopperle WM, DeWolf WC. The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. Stem Cells. 2007; 25:723-730. (Clone-specific). View Reference
  6. Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282:1145-1147. (Clone-specific: Immunocytochemistry (cytospins)). View Reference
  7. Thomson JA, Kalishman J, Golos TG, et al. Isolation of a primate embryonic stem cell line. Proc Natl Acad Sci U S A. 1995; 92:7844-7848. (Clone-specific: Immunocytochemistry (cytospins)). View Reference
View All (7) View Less
560193 Rev. 4

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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.