Confocal immunofluorescent analysis of NTERA-2 cells using TRA-1-81 (TRA-1-81) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).Learn more about how we get our images
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
posted June 2005
revised March 2016
Protocol Id: 293
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
Recommended Fluorochrome-conjugated Anti-Mouse secondary antibodies:
NOTE: When using any primary or fluorochrome-conjugated secondary antibody for the first time, titrate the antibody to determine which dilution allows for the strongest specific signal with the least background for your sample.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted December 2010
Protocol Id: 144
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: If live cell staining is desired, proceed to Section C.
NOTE: Count cells using a hemocytometer or alternative method.
posted January 2009
revised June 2017
Protocol Id: 36
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
TRA-1-81 detects endogenous levels of TRA-1-81 antigen.
Monoclonal antibody is produced by immunizing animals with human embryonal carcinoma 2102Ep cl.2A6 cells.
TRA-1-60 and TRA-1-81 antibodies detect antigens present on the surface of human stem, teratocarcinoma, and embryonic germ cells (1). TRA-1-60(S) reacts with a neuraminidase sensitive epitope of a proteoglycan (2,3), while TRA-1-81 reacts with a neuraminidase insensitive epitope on the same antigen. Recently this antigen has been proposed to be a form of the protein podocalyxin (4). TRA-1-60 is also detected in the serum of patients with germ cell tumors (5,6).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. DRAQ5 is a registered trademark of Biostatus Limited.
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|4745S||100 µl (200 sections)||$ 255.0|