Anti-TRF2 + TRF1 抗体 [TRF-78]

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

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.

This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

• ICC/IF

AbReview7136****

Immunocytochemistry/ Immunofluorescence - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

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.

This image is courtesy of an anonymous Abreview

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling TRF1 and TRF2 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-TRF2 + TRF1 antibody [TRF-78]. The antibody was developed using Goat Anti-Mouse IgG, Cy3 conjugate. Cells were counterstained with DAPI (blue) to stain nuclei.

• WB

Supplier Data

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

Anti-TERF antibody [TRF-78] (ab10579) staining at 4 ug/ml, shown in green. In Western blot, ab10579 was shown to bind specifically to TERF. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution.

All lanes:

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (ab10579) at 4 µg/mL

Lane 1:

Recombinant Human TRF1 protein cell lysate at 0.1 µg

Lane 2:

Recombinant Human TRF1 protein cell lysate at 0.5 µg

Lane 3:

Empty

Lane 4:

Recombinant Human TRF2 protein cell lysate at 0.1 µg

Lane 5:

Recombinant Human TRF2 protein cell lysate at 0.5 µg

Observed band size: 80 kDa,56 kDa

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• WB

AbReview67699****

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

Blocking step

Milk as blocking agent for 2 hours · Concentration : 5% · Temperature : 21°C.

Incubation time

12 hours · Temperature : 4°C · Diluent : 5% milk in TBST.

All lanes:

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (ab10579) at 1/1000 dilution

Lane 1:

SAOS2 (Human osteosarcoma cell line) cell lysate at 20 µg

Lane 2:

U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate at 20 µg

Secondary

All lanes:

Goat anti Mouse polyclonal IRDye 800CW at 1/1000 dilution

Predicted band size: 50 kDa

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Exposure time: 5min

Verified customer

• WB

Supplier Data

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

All lanes:

Western blot - Anti-TRF2 + 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

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• WB

CiteAb

Western blot - Anti-TRF2 + TRF1 antibody [TRF-78] (AB10579)

TRF2 + TRF1 western blot using anti-TRF2 + TRF1 antibody [TRF-78] ab10579. Publication image and figure legend from Wang, H., Zhang, K., et al., 2017, BMC Biol, PubMed 29216888.

ab10579 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab10579 please see the product overview.

TERF1 expression levels associated with telomere length and pluripotency of human embryonic stem cell (hESCs). a Scheme for hTERF1 knockout by CRISPR/Cas9. b Morphology of TERF1+/- and wild-type (WT) hESCs (WA26) at passage 11. Scale bar = 100 μm. c Expression levels of pluripotency and telomerase genes in TERF1+/- and WT hESCs by qPCR analysis. *p < 0.05, **p < 0.01, compared with WT controls. The data values of each gene are provided in Additional file 18 : Table S10. d Western blot analysis of TERF1, SOX2, and OCT4/POU5F1 protein levels in TERF1+/- and WT hESCs. β-actin served as a loading control. e Cell cycle analysis of TERF1+/- and WT hESCs by flow cytometry. Data represent mean ± SD of three independent experiments. f Telomerase activity by TRAP assay of TERF1+/- and WT hESCs at different passages. U2OS served as a negative control. g Telomere length measurement by southern blot analysis of TERF1+/- and WT hESCs. h Quantification of southern blot results. Data represent mean ± SD of two independent experiments. i Immunofluorescence detection of γH2AX and telomere FISH of TERF1+/- and WT hESCs. j, k Number of telomere-γH2AX colocalization foci per cell (j) and percentage of cells with the colocalization (k). l Relative telomere length of individual TERF1+/- and WT hESCs by scT&R-seq. Right panel shows the mean telomere length by scT&R-seq. m Gene expression measured in single cells of TERF1+/- and WT hESCs by scT&R-seq. Right panel shows the mean level of gene expression. n Linear regression analysis of telomere length and gene expression (TERF1 and NANOG) at single cell level

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