Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling TRF1 with ab10579. Cells were fixed and permeabilized with 4% paraformaldehyde followed by 0.5% Triton^™ X-100. Fixed cells were stained with 10 μg/mL Anti-TRF1 antibody [TRF-78] (ab10579).  The antibody was developed using Goat Anti-Mouse IgG, Cy3 conjugate. Cells were counterstained with DAPI (blue) to stain nuclei.

All lanes : Anti-TRF1 antibody [TRF-78] (ab10579) at 1/1000 dilution

Lane 1 : SAOS2 (Human osteosarcoma cell line) cell lysate
Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti Mouse polyclonal IRDye 800CW at 1/1000 dilution

Predicted band size: 50 kDa


Exposure time: 5 minutes

All lanes : Anti-TRF1 antibody [TRF-78] (ab10579) at 4 µg/ml

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 4 : U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate

Secondary
All lanes : Goat Anti-Mouse IgG-Peroxidase

Predicted band size: 50 kDa

Immunofluorescent imaging of human cells (U2OS) with ab10579 confirms the specificity of this antibody.  A few intense nuclear foci are seen in interphase cells, corresponding to telomeric localisation.  The complete absence of background nuclear or cytoplasmic staining confirms the specificity of this antibody. This image is in exact agreement with numerous published reports.

IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees. Nuclei are visualised using Hoechst stain. 

ab10579 at 1/500 staining human HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by ICC/IF. The cells were parafomaldehyde fixed and blocked with BSA prior to incubation with the antibody for 45 minutes. An Alexa Fluor^® 555 conjugated donkey anti-mouse antibody was used as the secondary.

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