711084

antibody from Invitrogen Antibodies

Targeting: TERT EST2, hEST2, TCS1, TP2, TRT

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711084

Provider

Invitrogen Antibodies

Product name

TERT Recombinant Polyclonal Antibody (19HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig, Dog and Bovine

Antibody clone number

19HCLC

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Nuclear enriched extracts (30 µg lysate) of HepG2 (Lane 1), A549 (Lane 2), Jurkat (Lane 3) and MCF-7 (Lane 4) and tissue extracts of Rat Brain (Lane 5). The blots were probed with Anti-TERT Recombinant Rabbit Polyclonal Antibody (Product # 711084, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 127 kDa and 120 kDa band corresponding to isoforms of TERT was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, Hep G2 cells were fixed and permeabilized for detection of endogenous Telomerase reverse transcriptase using Anti- TERT Recombinant Rabbit Polyclonal Antibody (Product # 711084, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of TERT protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of panels a, b and c clearly demonstrating nuclear localization of TERT. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.