TERT antibody (C-Term)

Catalog No. ABIN6655270

Product Details anti-TERT Antibody

Target: TERT (Telomerase Reverse Transcriptase (TERT))

Binding Specificity: C-Term

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This TERT antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Immunoprecipitation (IP), Fluorescence Microscopy (FM)

Purification: Affinity Purified Anti-hTERT Antibody was prepared from monospecific antiserum by immunoaffinity chromatography using synthetic peptide coupled to agarose beads. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum. Although it has been reported that this antibody reacts with mouse TERT (mTERT) (see Drissi, et al. 2001), the binding to mTERT is considerably weaker and less specific than the binding to hTERT (not shown).

Immunogen:

Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to a region near the carboxy terminal end of hTERT (accession number AF018167).

Immunogen Type: Peptide

Isotype: IgG

Application Details

Application Notes:

Immunohistochemistry Dilution: 1:500

Application Note: Anti-Telomerase catalytic subunit antibody has been tested for use in immunoblotting, immunoprecipitation, and immunofluorescence microscopy. In these assays, the antibody detects ectopically-expressed hTERT and high levels of endogenous hTERT. A SY5Y cell nuclear extract can be used as a positive control. This antibody primarily detects hTERT, but several non-specific bands appear on immunoblots. In immunofluorescence microscopy assays, staining with anti-TERT-16 was specific to the nuclei of cells with ectopic TERT expression. In immunoblot assays, whole cell or nuclear extracts were loaded at a concentration of 100 μg protein per well. A working dilution of 1:500 anti-TERT antibody was used followed by a 1:3,000 dilution of HRP goat anti-rabbit IgG as the secondary antibody. For immunofluorescence microscopy staining, a working dilution of 1:500 was used followed by a 1:200 dilution of rhodamine-conjugated donkey anti-rabbit IgG as a secondary antibody. Immunoprecipitation was performed using 20 μL of protein A beads and 2 μL of the anti-TERT serum per 1 mg protein from cell lysate. A working dilution of 1:500 is also suggested for immunohistochemistry. To detect TERT, fix cells in 2 % paraformaldehyde (in PBS) for 10'. Wash the slides twice in PBS for 5' each. Permeabilize the cells in 0.5 % NP-40 for 10'. Wash as before in PBS. Block the cells using PBG buffer (0.2 % cold water fish gelatin (Sigma G-7765) and 0.5 % BSA in PBS) for 20' at room temperature. Incubate in primary antibody (diluted in PBG) for 1-2 hours at RT or overnight at 4 °C. Wash the slides three times in PBG for 5' 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' each. Mount in DAPI-containing medium.

Immunoprecipitation Dilution: 2 μL per mg of lysate

ELISA Dilution: 1:10,000 - 1:50,000

Western Blot Dilution: 1:500- 1:2,000

IF Microscopy Dilution: 1:500

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

0.01 % (w/v) Sodium Azide

Stabilizer: None

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: RT,4 °C,-20 °C

Storage Comment: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

References for anti-TERT antibody (ABIN6655270)

Altamura, Martano, Licenziato, Maiolino, Borzacchiello et al.: "Telomerase Reverse Transcriptase (TERT) Expression, Telomerase Activity, and Expression of Matrix Metalloproteinases (MMP)-1/-2/-9 in Feline Oral Squamous Cell Carcinoma Cell Lines Associated With ..." in: Frontiers in veterinary science, Vol. 7, pp. 148, (2020) (PubMed).

Hugdahl, Kalvenes, Mannelqvist, Ladstein, Akslen: "Prognostic impact and concordance of TERT promoter mutation and protein expression in matched primary and metastatic cutaneous melanoma." in: British journal of cancer, Vol. 118, Issue 1, pp. 98-105, (2019) (PubMed).

Schmidt, Zaug, Kufer, Cech: "Dynamics of human telomerase recruitment depend on template- telomere base pairing." in: Molecular biology of the cell, (2018) (PubMed).

Kabacik, Horvath, Cohen, Raj: "Epigenetic ageing is distinct from senescence-mediated ageing and is not prevented by telomerase expression." in: Aging, Vol. 10, Issue 10, pp. 2800-2815, (2018) (PubMed).

Vanderstraeten, Tuyaerts, Everaert, Van Bree, Verbist, Luyten, Amant: "In Vitro Assessment of the Expression and T Cell Immunogenicity of the Tumor-Associated Antigens BORIS, MUC1, hTERT, MAGE-A3 and Sp17 in Uterine Cancer." in: International journal of molecular sciences, Vol. 17, Issue 9, (2017) (PubMed).

Li, Reddy, Comai: "The Werner Syndrome Helicase Coordinates Sequential Strand Displacement and FEN1-Mediated Flap Cleavage during Polymerase δ Elongation." in: Molecular and cellular biology, Vol. 37, Issue 3, (2017) (PubMed).

Hong, Lee, Chung: "Telomerase activates transcription of cyclin D1 gene through an interaction with NOL1." in: Journal of cell science, Vol. 129, Issue 8, pp. 1566-79, (2016) (PubMed).

Jeong, Kim, Lee, Cha, Khadka, Cho, Chung: "Akt-mediated phosphorylation increases the binding affinity of hTERT for importin α to promote nuclear translocation." in: Journal of cell science, Vol. 128, Issue 12, pp. 2287-301, (2015) (PubMed).

Rubis, Holysz, Gladych, Toton, Paszel, Lisiak, Kaczmarek, Hofmann, Rybczynska: "Telomerase downregulation induces proapoptotic genes expression and initializes breast cancer cells apoptosis followed by DNA fragmentation in a cell type dependent manner." in: Molecular biology reports, Vol. 40, Issue 8, pp. 4995-5004, (2014) (PubMed).

Radan, Hughes, Teichroeb, Vieira Zamora, Jewer, Postovit, Betts: "Microenvironmental regulation of telomerase isoforms in human embryonic stem cells." in: Stem cells and development, Vol. 23, Issue 17, pp. 2046-66, (2014) (PubMed).

Samba-Louaka, Stavru, Cossart: "Role for telomerase in Listeria monocytogenes infection." in: Infection and immunity, Vol. 80, Issue 12, pp. 4257-63, (2013) (PubMed).

Ponsot, Echaniz-Laguna, Delis, Kadi: "Telomere length and regulatory proteins in human skeletal muscle with and without ongoing regenerative cycles." in: Experimental physiology, Vol. 97, Issue 6, pp. 774-84, (2013) (PubMed).

Lee, Koh, Kim, Ahn, Kim: "Upregulation of Nrf2 expression by human cytomegalovirus infection protects host cells from oxidative stress." in: The Journal of general virology, Vol. 94, Issue Pt 7, pp. 1658-68, (2013) (PubMed).

Eitan, Tichon, Gazit, Gitler, Slavin, Priel: "Novel telomerase-increasing compound in mouse brain delays the onset of amyotrophic lateral sclerosis." in: EMBO molecular medicine, Vol. 4, Issue 4, pp. 313-29, (2012) (PubMed).

Gizard, Heywood, Findeisen, Zhao, Jones, Cudejko, Post, Staels, Bruemmer: "Telomerase activation in atherosclerosis and induction of telomerase reverse transcriptase expression by inflammatory stimuli in macrophages." in: Arteriosclerosis, thrombosis, and vascular biology, Vol. 31, Issue 2, pp. 245-52, (2011) (PubMed).

Georgopoulos, Kirkwood, Varley, MacLaine, Aziz, Southgate: "Immortalisation of normal human urothelial cells compromises differentiation capacity." in: European urology, Vol. 60, Issue 1, pp. 141-9, (2011) (PubMed).

Abreu, Aritonovska, Reichenbach, Cristofari, Culp, Terns, Lingner, Terns: "TIN2-tethered TPP1 recruits human telomerase to telomeres in vivo." in: Molecular and cellular biology, Vol. 30, Issue 12, pp. 2971-82, (2010) (PubMed).

Oh, Ghim, Lee, Yang, Kim, Ahn, Song: "PML-IV functions as a negative regulator of telomerase by interacting with TERT." in: Journal of cell science, Vol. 122, Issue Pt 15, pp. 2613-22, (2009) (PubMed).

Ahmed, Passos, Birket, Beckmann, Brings, Peters, Birch-Machin, von Zglinicki, Saretzki: "Telomerase does not counteract telomere shortening but protects mitochondrial function under oxidative stress." in: Journal of cell science, Vol. 121, Issue Pt 7, pp. 1046-53, (2008) (PubMed).

Wu, Dudognon, Nguyen, Hillion, Pendino, Tarkanyi, Aradi, Lanotte, Tong, Chen, Ségal-Bendirdjian: "Immunodetection of human telomerase reverse-transcriptase (hTERT) re-appraised: nucleolin and telomerase cross paths." in: Journal of cell science, Vol. 119, Issue Pt 13, pp. 2797-806, (2006) (PubMed).

Target Details for TERT

Target: TERT (Telomerase Reverse Transcriptase (TERT))

Alternative Name: Telomerase catalytic subunit (TERT Products)

Synonyms: CMM9 antibody, DKCA2 antibody, DKCB4 antibody, EST2 antibody, PFBMFT1 antibody, TCS1 antibody, TP2 antibody, TRT antibody, hEST2 antibody, hTRT antibody, chTERT antibody, Tert antibody, ATTERT antibody, F5E19.190 antibody, F5E19_190 antibody, telomerase reverse transcriptase antibody, tert antibody, TR antibody, est2 antibody, tcs1 antibody, telomerase antibody, tert-A antibody, tp2 antibody, trt antibody, xTERT antibody, telomerase reverse transcriptase antibody, Telomerase Reverse Transcriptase antibody, Tertp antibody, Telomerase catalytic subunit antibody, telomerase reverse transcriptase L homeolog antibody, TERT antibody, trt-1 antibody, Tert antibody, tert antibody, CMU_003490 antibody, VDBG_04487 antibody, GL50803_16225 antibody, tert.L antibody

Background:

Synonyms: rabbit anti-TERT antibody, rabbit anti-Telomerase catalytic subunit antibody, hTERT, Telomerase reverse transcriptase, HEST2, Telomerase-associated protein 2, TP2, EST2, TCS1, TRT

Background: 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.

Gene Name: TERT

Gene ID: 7015

UniProt: O14746

Pathways: Telomere Maintenance