Anti-Telomerase reverse transcriptase antibody [2C4] (ab5181)

Overview

  • Product nameAnti-Telomerase reverse transcriptase antibody [2C4]
    See all Telomerase reverse transcriptase primary antibodies
  • Description
    Mouse monoclonal [2C4] to Telomerase reverse transcriptase
  • Tested applicationsFlow Cyt, IHC-P, ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (Human) from insect cells.

  • Positive control
    • Immunofluorescence/immunocytochemistry: 293T, Hela, BJ, WI-38 Western Blot: 293T Whole Cell Lysates, Hela Whole Cell Lysates Immunohistochemistry-Paraffin: Pancreatic carcinoma, Normal pancreas (human)

Properties

Applications

Our Abpromise guarantee covers the use of ab5181 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1-2µg for 106 cells. (Also see Abreview)
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. Fix with 1:1 ice cold MEOH:acetone and place at -20°C for 10 minutes.
WB 1/500 - 1/1000. Detects a band of approximately 127 kDa (predicted molecular weight: 127 kDa).

An additional background band at ~110 kDa may be present. Please note the isotype for this product is IgM and requires the use of an IgM secondary. Even though we have a positive Abreview on IHC, this is not a batch tested application and we cannot guarantee that it will work.

IP Use at an assay dependent concentration.

Target

  • FunctionTelomerase is a ribonucleoprotein enzyme essential for the replication of chromosome termini in most eukaryotes. Active in progenitor and cancer cells. Inactive, or very low activity, in normal somatic cells. Catalytic component of the teleromerase holoenzyme complex whose main activity is the elongation of telomeres by acting as a reverse transcriptase that adds simple sequence repeats to chromosome ends by copying a template sequence within the RNA component of the enzyme. Catalyzes the RNA-dependent extension of 3'-chromosomal termini with the 6-nucleotide telomeric repeat unit, 5'-TTAGGG-3'. 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. More active on substrates containing 2 or 3 telomeric repeats. Telomerase activity is regulated by a number of factors including telomerase complex-associated proteins, chaperones and polypeptide modifiers. Modulates Wnt signaling. Plays important roles in aging and antiapoptosis.
  • Tissue specificityExpressed at a high level in thymocyte subpopulations, at an intermediate level in tonsil T lymphocytes, and at a low to undetectable level in peripheral blood T lymphocytes.
  • Involvement in diseaseNote=Activation of telomerase has been implicated in cell immortalization and cancer cell pathogenesis.
    Defects in TERT are associated with susceptibilty to aplastic anemia (AA) [MIM:609135]. AA is a rare disease in which the reduction of the circulating blood cells results from damage to the stem cell pool in bone marrow. In most patients, the stem cell lesion is caused by an autoimmune attack. T-lymphocytes, activated by an endogenous or exogenous, and most often unknown antigenic stimulus, secrete cytokines, including IFN-gamma, which would in turn be able to suppress hematopoiesis.
    Note=Genetic variations in TERT are associated with coronary artery disease (CAD).
    Defects in TERT are a cause of dyskeratosis congenita autosomal dominant (ADDKC) [MIM:127550]; also known as dyskeratosis congenita Scoggins type. ADDKC is a rare, progressive bone marrow failure syndrome characterized by the triad of reticulated skin hyperpigmentation, nail dystrophy, and mucosal leukoplakia. Early mortality is often associated with bone marrow failure, infections, fatal pulmonary complications, or malignancy.
    Defects in TERT are a cause of susceptibility to pulmonary fibrosis idiopathic (IPF) [MIM:178500]. Pulmonary fibrosis is a lung disease characterized by shortness of breath, radiographically evident diffuse pulmonary infiltrates, and varying degrees of inflammation and fibrosis on biopsy. It results in acute lung injury with subsequent scarring and endstage lung disease.
  • Sequence similaritiesBelongs to the reverse transcriptase family. Telomerase subfamily.
    Contains 1 reverse transcriptase domain.
  • DomainThe primer grip sequence in the RT domain is required for telomerase activity and for stable association with short telomeric primers.
    The RNA-interacting domain 1 (RD1)/N-terminal extension (NTE) is required for interaction with the pseudoknot-template domain of each of TERC dimers. It contains anchor sites that bind primer nucleotides upstream of the RNA-DNA hybrid and is thus an essential determinant of repeat addition processivity.
    The RNA-interacting domain 2 (RD2) is essential for both interaction with the CR4-CR5 domain of TERC and for DNA sythesis.
  • Post-translational
    modifications
    Ubiquitinated, leading to proteasomal degradation.
    Phosphorylation at Tyr-707 under oxidative stress leads to translocation of TERT to the cytoplasm and reduces its antiapoptotic activity. Dephosphorylated by SHP2/PTPN11 leading to nuclear retention. Phosphorylation by the AKT pathway promotes nuclear location.
  • Cellular localizationNucleus > nucleolus. Nucleus > nucleoplasm. Nucleus. Chromosome > telomere. Cytoplasm. Nucleus > PML body. Shuttling between nuclear and cytoplasm depends on cell cycle, phosphorylation states, transformation and DNA damage. Diffuse localization in the nucleoplasm. Enriched in nucleoli of certain cell types. Translocated to the cytoplasm via nuclear pores in a CRM1/RAN-dependent manner involving oxidative stress-mediated phosphorylation at Tyr-707. Dephosphorylation at this site by SHP2 retains TERT in the nucleus. Translocated to the nucleus by phosphorylation by AKT.
  • Information by UniProt
  • Database links
  • Alternative names
    • CMM9 antibody
    • DKCA2 antibody
    • DKCB4 antibody
    • EST2 antibody
    • hEST2 antibody
    • htert antibody
    • hTRT antibody
    • PFBMFT1 antibody
    • TCS1 antibody
    • Telomerase associated protein 2 antibody
    • Telomerase Catalytic Subunit antibody
    • Telomerase reverse transcriptase antibody
    • Telomerase-associated protein 2 antibody
    • Telomere Reverse Transcriptase antibody
    • TERT antibody
    • TERT_HUMAN antibody
    • TP2 antibody
    • TRT antibody
    see all

Anti-Telomerase reverse transcriptase antibody [2C4] images



  • Predicted band size : 127 kDa


    ab5181 at a 1/500 dilution staining approximately 127kDa Telomerase in MJ90 cell lysate by Western blot (ECL).
  • Overlay histogram showing Jurkat cells stained with ab5181 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5181, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Telomerase reverse transcriptase antibody [2C4] (ab5181)

This product has been referenced in:
  • Friis-Ottessen M  et al. Reduced hTERT protein levels are associated with DNA aneuploidy in the colonic mucosa of patients suffering from longstanding ulcerative colitis. Int J Mol Med 33:1477-83 (2014). IHC-P ; Human . Read more (PubMed: 24676865) »
  • Hemmaid KZ  et al. Impact of Hepatitis C Virus (HCV) infection on biomolecular markers influencing the pathogenesis of bladder cancer. Infect Agent Cancer 8:24 (2013). IHC ; Human . Read more (PubMed: 23809295) »

See all 12 Publications for this product

Product Wall

Application Western blot
Loading amount 25000 cells
Gel Running Conditions Non-reduced Non-Denaturing (Native) (8)
Sample Human Cell lysate - whole cell (prostate)
Specification prostate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Aug 04 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (Hepatocyte)
Specification Hepatocyte
Permeabilization No
Fixative 1:1 ice cold MEOH:acetone
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Submitted Feb 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 25 µg
Gel Running Conditions Non-reduced Denaturing (15%)
Sample Human Cell lysate - whole cell (Hep3B)
Specification Hep3B
Treatment 1ug/ml doxorubicin 24h
Blocking step I-Block(Applied biosystems) as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
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Submitted Jul 08 2013

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I am very pleased to hear you would like to accept our offer and test ab5181 in Mouse. This code will give you 1 freeprimary antibodybefore the expiration date (ab104588, ab94902 or any oth...

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Thank you very much for your interest in ab5181.

Unfortunately, because we carry over 75,000 products, it isn't feasible for us to keep small sample sizes of our products.

To our knowledge, ab5181 has not been tested in Mouse and the ...

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Thank you for contacting us again.

We usually, if not mentioned on the datasheet, use the protein sequencesas described in SwissProt (ID#O14746). This database, however, has a link to NCBI Sequence databases as a cross- reference.

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Thank you for contacting us.

I would like to suggestthe followingtwo antibodies which are suitable for your customer's purpose:

http://www.abcam.com/index.html?datasheet=5181 (or use the following: http://www.abcam.com/index.html?d...

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Thank you for contacting us.

There isn’t a specific protocol for performing IHC-P using this antibody. According to the datasheet the antibody concentration is assay dependant, and optimisation is recommended by using different antibody...

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Thank you for your reply and I understand your customer's concern. We were supplying this antibody from an outside lab and were selling it at 100ul and it was unpurified so the concentration was not determined. Now we currently make this clone in house...

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Thank you for your reply. The concentration for all lots of this antibody (GR59361-1 and GR22704-2) have not been determined because it is a concentrated tissue culture supernatant. I apologize for the misinformation on the datasheet. I am having our D...

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