Telomerase (EST2, hEST2, TCS1, Telomerase Catalytic Subunit, Telomerase-associated protein 2, Telomere Reverse Transcriptase, TERT, TP2, TRT)

Telomerase is a reverse transcriptase that adds telomeric repeats (TTAGGG)n to chromosomal ends, compensating for the telomere shortening that occurs with DNA replication. In normal human somatic cells, telomerase is repressed and telomeres progressively shorten, leading to limited lifespan and senescence. Reactivation of telomerase activity is associated with human cancer and cell immortalization. Approximately 85% of human cancers, including breast, prostate, stomach, bladder, colon, and liver cancer, have telomerase activity, whereas most normal somatic cells do not. The specificity of telomerase to human cancer has led to investigations of telomerase activity and expression as a tumor marker. For example, the presence of telomerase activity in human urine has been identified as a marker for human bladder carcinoma. Human telomerase consists of three major subunits: a catalytic protein subunit called hTERT (for human TElomerase Reverse Transcriptase), a template RNA called hTR, and telomerase-associated protein (TEP-1). TERT and hTR are minimally required to reconstitute telomerase activity in vitro. In human cells, hTR is constitutively expressed; TERT transcription is a primary mechanism for regulation of telomerase activity.\nas a positive control.\n\nApplications: \nSuitable for use in ELISA, Immunofluorescence, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.\n\nRecommended Dilution:\nELISA: 1:10,000-1:50,000\nImmunofluorescence: 1:500, followed by a 1:200 dilution of rhodamine conjugated donkey anti-rabbit IgG as secondary antibody.\nWestern Blot: 1:500-1:1000 using a 1:3000 dilution of HRP conjugated goat anti-rabbit IgG as secondary antibody. Whole cell or nuclear extracts were loaded at a concentration of 100ug protein/well.\nImmunoprecipitation: 2ul/20ul of protein A beads/1mg protein from cell lysate.\nImmunohistochemistry (Fixed cells): 1:500. Fix cells in PBS, 2% paraformaldehyde for 10 minutes. Wash the slides twice in PBS for 5 minutes each. Permeabilize the cells in 0.5% NP-40 for 10 minutes. Wash as before in PBS. Block the cells using PBG buffer (0.2% cold water fish gelatin, 0.5% BSA in PBS) for 20 minutes at RT. Incubate in T2399-29 (diluted in PBG) for 1-2 hours at RT or overnight at 4C. Wash the slides three times in PBG for 5 minutes each. Incubate with secondary antibody (diluted in PBG) for 1 hour at RT in the dark. Wash the slides three times in PBG for 5 minutes each. Mount in DAPI-containing medium.\nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nSY5Y cell nuclear extract.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

T2399-29

Size

100ug

Applications

ELISA, IF, IHC, IP, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS, pH 7.2, 0.1% sodium azide. No stabilizing proteins added.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1.Drissi, R., Zindy, F., Roussel, M. F., and Cleveland, J.L. (2001) c-MYC-mediated regulation of telomerase activity is disabled in immortalized cells. J Biol Chem 276:32, 29994-30001. 2.Shay JW, Zou Y, Hiyama E, Wright WE. (2001) . Hum Mol Genet 2001; 10:677-85.3. Hiyama E, Hiyama K, Yokoyama T, Shay JW. (2001) . Neoplasia 2001; 3:17-26.4. Wick M, Zubov D, Hagen G. (1999) Genomic organization and promoter characterization of the gene encoding the human telomerase reversetranscriptase (hTERT). Gene 1999; 232:97-106.

Additional Info

Recognizes human Telomerase, but several non-specific bands appear on Western Blot. Reacts with ectopically-expressed human Telomerase and high levels of endogenous human Telomerase.