Podocalyxin-Like (PODXL) antibody

Details for Product No. ABIN967660
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Synonyms PODXL, podxl, fc18d02, wu:fc18d02, Gp200, PC, PCLP, PCLP-1, AW121214, Ly102, Pclp1, podocalyxin, Gp135, PCLP1, cPCLP1, MEP21, Podxl
(112), (15), (9), (1)
(54), (51), (15), (8), (2)
Clonality (Clone)
Monoclonal ()
(11), (11), (7), (5), (3), (3), (2), (2), (2), (1)
Western Blotting (WB), BioImaging (BI), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunoprecipitation (IP), Radioimmunoassay (RIA)
(105), (46), (43), (21), (15), (13), (10), (10), (5), (3), (3), (2), (1), (1), (1)
Pubmed 7 references available
Quantity 0.1 mg
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Catalog No. ABIN967660
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Immunogen Human Embryonal Carcinoma Cell Line
Clone TRA-1-81
Isotype IgM, kappa
Cross-Reactivity Rhesus Monkey
Characteristics 1. Please refer to us for technical protocols.
2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name TRA-1-81
Background The TRA-1-81 monoclonal antibody reacts with a pluripotent-stem-cell-specific epitope on a high-molecular-weight transmembrane glycoprotein. The TRA-1-81 antigen is an epitope on the same keratan sulfate core molecule, podocalyxin, as 4 other distinct antigens on tumor-derived cell lines, TRA-1-60, GCTM2, K4, and K21. The expression of TRA-1-81 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. As human EC and ES cells undergo differentiation, expression of TRA-1-81 antigen is lost.
Molecular Weight multiple, >200 kDa
Application Notes Bioimaging
1. Seed the cells in appropriate culture medium at an appropriate cell density in an 96-well Imaging Plate , and culture overnight to 48 hours.
2. Remove the culture medium from the wells, wash the wells twice with 100 myl of 1× PBS, and fix the cells by adding 100 µl of fresh 3.7% Formaldehyde in PBS or fixation buffer to each well and incubating for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and wash the wells twice with 100 myl of 1× PBS.
4. Dilute the antibody in 1× PBS, and stain the cells by adding 50 µl of the diluted antibody to each well and incubating for 1 hour at RT.
5. Remove the diluted antibody, and wash the wells three times with 100 myl of 1× PBS.
6. Remove the PBS, dilute the second-step reagent in 1× PBS, and stain the cells by adding 50 µl of the diluted second-step reagent to each well and incubating for 1 hour at RT.
7. Remove the diluted second-step reagent, and wash the wells twice with 100 myl of 1× PBS.
8. Remove the PBS, and counter-stain the nuclei by adding 100 ml of a 2 mg/ml solution of Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
9. View and analyze the cells on an appropriate imaging instrument.

Related Products: ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 0.5 mg/ml
Buffer Aqueous buffered solution.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C
Supplier Images
anti-Podocalyxin-Like (PODXL) antibody Immunofluorescent staining of human ES cell line. The H9 cell line (WiCell, Madison, WI) was cultured, fixed, and stained with Purified Mouse anti-Human TRA-1-81 Antigen monoclonal antibody (pseudo-colored green) according to the Recommended Assay Procedure. The second-step reagent was Alexa Fluor® 647 goat anti-mouse Ig (Invitrogen). The left image shows the plasma membrane staining by the TRA-1-81 mAb, and the right image shows TRA-1-81 with counter-staining of the nuclei by Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer using a 10X objective and merged using BD Attovision™ software.
anti-Podocalyxin-Like (PODXL) antibody (2) Immunofluorescent staining of human ES cell line
anti-Podocalyxin-Like (PODXL) antibody (3) Western Blot analysis of TRA-1-60 in mouse and human ES cell lines. Lysates from ES-E14TG2a mouse ES cells (ATCC CRL-1821, left blot) and H9 human ES cells (WiCell, Madison, WI, right blot) were probed with purified mouse anti-human TRA-1-60 Antigen monoclonal antibody at titrations of 2.0 (lanes 1), 1.0 (lanes 2), and 0.5 µg/ml (lanes 3). High-molecular-weight molecules bearing the TRA-1-60 epitope are identified above 200 kDa in the human ES cells. Expression of the TRA-1-60 epitope has not been reported in mouse ES cells.
The H9 cells were cultured on a mitomycin C-treated mouse embryonic fibroblast feeder layer [MEF (CF-1), ATCC SCRC-1040] that maintains the undifferentiated state of the ES cells. The lysate was made from a mixture of the 2 cell types, the majority of which were H9 cells.
Product cited in: Schopperle, DeWolf: "The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma." in: Stem cells (Dayton, Ohio), Vol. 25, Issue 3, pp. 723-30, 2007 (PubMed).

Henderson, Draper, Baillie et al.: "Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens." in: Stem cells (Dayton, Ohio), Vol. 20, Issue 4, pp. 329-37, 2002 (PubMed).

Draper, Pigott, Thomson et al.: "Surface antigens of human embryonic stem cells: changes upon differentiation in culture." in: Journal of anatomy, Vol. 200, Issue Pt 3, pp. 249-58, 2002 (PubMed).

Badcock, Pigott, Goepel et al.: "The human embryonal carcinoma marker antigen TRA-1-60 is a sialylated keratan sulfate proteoglycan." in: Cancer research, Vol. 59, Issue 18, pp. 4715-9, 1999 (PubMed).

Thomson, Itskovitz-Eldor, Shapiro et al.: "Embryonic stem cell lines derived from human blastocysts." in: Science (New York, N.Y.), Vol. 282, Issue 5391, pp. 1145-7, 1998 (PubMed).

Thomson, Kalishman, Golos et al.: "Isolation of a primate embryonic stem cell line." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, Issue 17, pp. 7844-8, 1995 (PubMed).

Andrews, Banting, Damjanov et al.: "Three monoclonal antibodies defining distinct differentiation antigens associated with different high molecular weight polypeptides on the surface of human embryonal carcinoma cells." in: Hybridoma, Vol. 3, Issue 4, pp. 347-61, 1985 (PubMed).

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