Technical Data
T2399-05
Telomerase (EST2, hEST2, TCS1, Telomerase Catalytic Subunit, Telomerase-associated protein 2, Telomere Reverse Transcriptase, TERT, TP2, TRT)
Description:
The 3'ends of chromosomes are capped with telomere sequences (TTAGGG; 6-26 nucleotides in length) by ribonucleoprotein telomerase during DNA replication. Telomerase is an unusual RNA-dependent DNA polymerase that uses and RNA component to specify the addition of telomere. The telomeric RNA contains a sequence complementary to TTAGGG. In ciliated protozoa and yeast, telomere length is maintained by regulating the activity of telomerase. Many mammalian cells do not express telomerase resulting into shortening of telomere with each cell division, and ultimately causing the chromosomal instability, aging and cell death. Approx. 4.8kb of telomeric DNA is lost with each cell division resulting into large number of chromosomal abnormalities. Most recently, introduction of telomerase into normal human cells has been shown to extend normal cell life by ~20 doubling. (2).

Purification of telomerase from the ciliate also revealed two protein of 43 and 123kD. p123 is a homolog of yeast Est2 (Essential for Telomerase activity). Mammalian homologs of yeast Est2 (also known as TP2 for Telomerase associated Protein 2; hEST2 or telomerase catalytic subunit or telomerase reverse transcriptase, TERT) have also been cloned (human Est2, 1132aa; mouse Est2 1122aa; ~127kD). hEST2 was localized in the nucleus. Yeast telomerase is 988aa protein.

Applications:
Suitable for use in ELISA, Western Blotting. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure using ECL. A band of 95-110kD has been detected in brush border membrane (2).
ELISA: 1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity purified. Control peptide can be used to coat ELISA plates at 1ug/ml.

Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Sold under license from Geron
TypeIsotypeCloneGrade
PabIgYSerum
SizeStorageShippingSourceHost
100ul-20CBlue IceYeastChicken
Concentration:
As reported
Immunogen:
A 23 AA Peptide sequence mapping near the N-terminus of yeast telomerase/Est2 (1). The peptide was coupled to KLH
Purity:
Serum
Form
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity:
The 23 AA yeast control peptide has no significant sequence homology with mammalian telomerases. We recommend the use EST21-23 for detection of human and mouse telomerase
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Nakamura Tm et al (1997) Science 277, 955-959;
2. Meyerson M et al (1997) Cell 90, 785-795; Greenberg RA et al (1998) Oncogene in press; Nugent CI et al (1998) Genes Develop. 12, 1073-85 (review).