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.
4.8 kb 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.
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, 1132 aa; mouse Est2 1122 aa; ~127kD).
hEST2 was localized in the nucleus.
A 16aa Peptide (EST21-P; Control peptide) sequence mapping in the middle of human Est2 (1) was selected for antibody production.
The peptide was coupled to KLH, and antibodies generated in rabbits.
Antibody has been affinity purified using control peptide-Sepharose.
Western blot positive control are prepared from human tumor cells (MDA231 breast cancer) expressing relatively high level of Est2.
WB Controls are provided in sample buffer and should be loaded 10-20/lane to visualize hEst2.
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 antibody using ECL.
The antibody has detected ~120-130kD band from several tumor cell lines.
An antibody made to a region near to the hESt21 epitope recognized ~125kD band in human and mouse tissues (2).
ELISA: 1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity pure.
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 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°C.
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
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Serum
Purity：Serum
Form：Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity：The 16 AA human control peptide showed 62% (10/16 aa) sequence homology with the mouse Est2. No significant sequence homology is detected with any other protein. We recommend the use EST23 (antibody # 3) for detection of mouse Est2
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu
Immunogen：A 16 AA Peptide sequence mapping in the middle of human Est2 (1). The peptide was coupled to KLH
