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
as a positive control.
Applications:Suitable for use in ELISA, Immunofluorescence, Western Blot, Immunoprecipitation and Immunohistochemistry.
Other applications not tested.
Recommended Dilution:ELISA: 1:10,000-1:50,000Immunofluorescence: 1:500, followed by a 1:200 dilution of rhodamine conjugated donkey anti-rabbit IgG as secondary antibody
Western 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
Immunoprecipitation: 2ul/20ul of protein A beads/1mg protein from cell lysate
Immunohistochemistry (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 4ºC.
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
Optimal dilutions to be determined by the researcher.
Positive Control:SY5Y cell nuclear extract.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°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.
仕様
Size：100ug
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide. No stabilizing proteins added.
Specificity：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.
Isotype：IgG
Calc Applications Abbrev：E IF IHC IP WB
Calc Crossreactivity：Hu
Immunogen：Synthetic peptide corresponding to a region near the C-terminal end of human Telomerase (Accession# AF018167).
