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Key Documents

MABC1795

Sigma-Aldrich

Anti-phospho-TERT (Thr249) Antibody, clone TpMab-3

Synonym(s):

EC:2.7.7.49, HEST2, TP2, Telomerase catalytic subunit, Telomerase reverse transcriptase, Telomerase-associated protein 2

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

TpMab-3, monoclonal

mol wt

calculated mol wt 127 kDa
observed mol wt ~127 kDa

purified by

using protein G

species reactivity

human

packaging

antibody small pack of 100

technique(s)

immunofluorescence: suitable
immunohistochemistry: suitable
western blot: suitable

isotype

IgG1κ

epitope sequence

N-terminal half

Protein ID accession no.

UniProt accession no.

storage temp.

2-8°C

Gene Information

human ... TERT(7015)

Specificity

Clone TpMab-3 is a mouse monoclonal antibody that detects phospho-TERT (Thr249).

Immunogen

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphothreonine 249 from the N-terminal half of human Telomerase reverse transcriptase (TERT).

Application

Quality Control Testing

Isotype testing: Identity confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be mouse IgG1k.

Tested Applications

Immunofluorescence Analysis: A representative lot detected TERT in Immunofluorescence applications (Matsuda, Y., et al. (2022). J Pathol. 257(2):172-185).

Western Blotting Analysis: A 1:250 dilution from a representative lot detected TERT in Mitotic HeLa cells with RO3306 treatment (negative) versus mitotic HeLa cell samples (positive), where the positive control samples were obtained by IP using clone 10E9-2 to enrich hTERT ( Data courtesy of Dr. Kenkichi Masutomi, National Cancer Center Research Institute, Tokyo, Japan).

Western Blotting Analysis: A representative lot detected TERT in Western Blotting applications (Matsuda, Y., et al. (2022). J Pathol. 257(2):172-185).

Immunohistochemistry Applications: A representative lot detected TERT in Immunohistochemistry applications (Matsuda, Y., et al. (2022). J Pathol. 257(2):172-185).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Telomerase reverse transcriptase (UniProt: O14746; also known as EC:2.7.7.49, HEST2, Telomerase catalytic subunit, Telomerase-associated protein 2, TP2) is encoded by the TERT (also known as EST2, TCS1, TRT) gene (Gene ID: 7015) in human. Telomerase is a ribonucleoprotein enzyme essential for the replication of chromosome termini in most eukaryotes. Telomerase reverse transcriptase (TERT) is the catalytic subunit of the telomerase responsible for adding TTAGGG repeats to the chromosome telomere ends. The catalytic cycle involves primer binding, primer extension and release of product once the template boundary has been reached or nascent product translocation followed by further extension. Cells with low or no telomerase expression lose telomere repeats during cell division, eventually resulting in cellular senescence. Most cancer cells, germ cells, and embryonic stem cells express high levels of telomerase, thus contributing to pluripotency and immortality. In addition to its telomere maintenance function, telomerase also has a pro-survival role in cellular resistance against DNA damage and ensuing apoptosis. Most cancer cells are highly proliferative and express high levels of nuclear telomerase activity. TERT is reported to shuttle between nuclear and cytoplasm and this shuttling is depended upon cell cycle, phosphorylation state, transformation, and DNA damage. TERT is also detected in the mitochondria where it helps prevent nuclear DNA damage by decreasing mitochondrial reactive oxygen species (ROS). Following exposure to H2O2 or -irradiation, cancer cells capable of excluding TERT from the nucleus display little or no DNA damage, while TERT nuclear retainment results in high DNA damage. Phosphorylation at tyrosine 707 under oxidative stress leads to translocation of TERT to the cytoplasm and reduces its anti-apoptotic activity and its dephosphorylation by SHP2/PTPN11 leads to its nuclear retention. Phosphorylation at serine 227 by AKT is reported to promote nuclear location. Phosphorylation of TERT at threonine 249 is reported to be a novel tumor biomarker of aggressive cancer with poor prognosis. (Ref.: Masuda, Y., et al. (2022). J. Pathol. 257(2); 172-185; Jacob, S., et al. (2008). J. Biol. Chem. 283(48); 33155-33161; Haendeler, J., et al. (2004). Circ. Res. 94(6); 768-775).

Physical form

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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