TRA1-60 antibody

Catalog No. ABIN967659

Product Details

Target: TRA1-60

Reactivity: Human, Rhesus Monkey

Host: Mouse

Clonality: Monoclonal

Conjugate: Un-conjugated

Application: Flow Cytometry (FACS), Immunoprecipitation (IP), Western Blotting (WB), BioImaging (BI)

Brand: BD Pharmingen™

Cross-Reactivity: Rhesus Monkey

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. 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.
3. Please refer to us for technical protocols.
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: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Embryonal Carcinoma Cell Line

Clone: TRA-1-60

Isotype: IgM kappa

Application Details

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.

Comment:

Related Products: ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

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

Storage Comment: Store undiluted at 4°C.

References

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).

Draper, Pigott, Thomson, Andrews: "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).

Henderson, Draper, Baillie, Fishel, Thomson, Moore, Andrews: "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).

Thomson, Itskovitz-Eldor, Shapiro, Waknitz, Swiergiel, Marshall, Jones: "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, Durning, Harris, Becker, Hearn: "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, Arnaud, Avner: "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).

Target Details

Target: TRA1-60

Alternative Name: TRA-1-60

Background: 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).
Synonyms: TRA-1-60(R)

Molecular Weight: Multiple, >200 kDa