TERT Antikörper (C-Term)

Produktnummer ABIN6655122

Produktdetails anti-TERT Antikörper

Target: TERT (Telomerase Reverse Transcriptase (TERT))

Bindungsspezifität: C-Term

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser TERT Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Immunofluorescence (IF), Immunoprecipitation (IP), Fluorescence Microscopy (FM)

Verwendungszweck: hTERT Antibody

Kreuzreaktivität (Details): Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Aufreinigung: Affinity Purified Anti-hTERT Antibody was prepared from monospecific antiserum by immunoaffinity chromatography using synthetic peptide coupled to agarose beads.

Sterilität: Sterile filtered

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).

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise:

Immunoprecipitation_Dilution: IP 2μL per mg lysate

ELISA_Dilution: 1:10,000 - 1:50,000

Immunohistochemistry_Dilution: 1:500

IF_Microscopy_Dilution: 1:500

Western_Blot_Dilution: 1:500

Kommentare:

Suggested Applications: ChIP, FISH, IHC, Multiplex
Anti-Telomerase catalytic subunit antibody has been tested for use in ELISA, 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 1mg 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Buffer:

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

Stabilizer: None

Preservative: 0.01 % (w/v) Sodium Azide

Konservierungsmittel: Sodium azide

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

Lagerung: -20 °C

Informationen zur Lagerung: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 μL). To minimize loss of volume dilute 1:10 by adding 225 μL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.

Haltbarkeit: 3 months

Referenzen für anti-TERT Antikörper (ABIN6655122)

Bernal, Zafon, Domínguez, Bertran, Tusell: "Generation of Immortalised But Unstable Cells after hTERT Introduction in Telomere-Compromised and p53-Deficient vHMECs." in: International journal of molecular sciences, Vol. 19, Issue 7, (2018) (PubMed).

Eitan, Braverman, Tichon, Gitler, Hutchison, Mattson, Priel: "Excitotoxic and Radiation Stress Increase TERT Levels in the Mitochondria and Cytosol of Cerebellar Purkinje Neurons." in: Cerebellum (London, England), Vol. 15, Issue 4, pp. 509-17, (2017) (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).

Theurillat, Udeshi, Errington, Svinkina, Baca, Pop, Wild, Blattner, Groner, Rubin, Moch, Privé, Carr, Garraway: "Prostate cancer. Ubiquitylome analysis identifies dysregulation of effector substrates in SPOP-mutant prostate cancer." in: Science (New York, N.Y.), Vol. 346, Issue 6205, pp. 85-9, (2014) (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).

Details zu TERT

Target: TERT (Telomerase Reverse Transcriptase (TERT))

Andere Bezeichnung: TERT (TERT Produkte)

Hintergrund:

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

Gen-ID: 7015

NCBI Accession: NP_001180305

UniProt: O14746

Pathways: Telomere Maintenance