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BV421 Mouse Anti-Human TRA-1-60 Antigen
BV421 Mouse Anti-Human TRA-1-60 Antigen
Flow cytometric analysis of TRA-1-60 expression of human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 50 grown on irradiated mouse embryonic fibroblasts were harvested with Accutase™ (Cat. No. 561527) and stained with either BD Horizon™ BV421 Mouse anti-TRA-1-60 (solid line) or BD Horizon™ BV421 Mouse IgM, κ isotype control (Clone G155-228, Cat. No.562704, dashed line). Flow cytometry was performed on a BD™ LSRII flow cytometry system.
Flow cytometric analysis of TRA-1-60 expression of human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 50 grown on irradiated mouse embryonic fibroblasts were harvested with Accutase™ (Cat. No. 561527) and stained with either BD Horizon™ BV421 Mouse anti-TRA-1-60 (solid line) or BD Horizon™ BV421 Mouse IgM, κ isotype control (Clone G155-228, Cat. No.562704, dashed line). Flow cytometry was performed on a BD™ LSRII flow cytometry system.
Product Details
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BD Horizon™
TRA-1-60(R)
Human (QC Testing)
Mouse BALB/c IgM, κ
Human Embryonal Carcinoma Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_2737738
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
  9. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
562711 Rev. 1
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).

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.

562711 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
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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). 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). 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). 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). 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). View Reference
View All (7) View Less
562711 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.