rabbit antibody against trf2  (Novus Biologicals)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Meiotic chromosome dynamics and double strand break formation in reptiles

doi: 10.3389/fcell.2022.1009776

Figure Lengend Snippet: Telomere dynamics during prophase I. Spermatocyte spreads labelled with antibodies against SYCP3 (green) and TRF2 (red), counterstaining the DNA with DAPI (blue) for (A) the bearded dragon, (B) the ocelot gecko and (C) the western banded gecko. Scale bar: 10 μm and 2 μm (insets). White arrowheads: telomeres from which SC is beginning to assemble. Yellow arrowheads: completely associated micro-chromosomes (i.e., lateral elements of the SC completely assembled between both telomeric ends).

Article Snippet: Immuno-staining of meiocytes was performed using the following primary antibodies: rabbit antibody against SYCP3 (#ab15093, Abcam, 1:100 dilution), rabbit antibody against SYCP1 (#ab15087, Abcam, 1:100 dilution), rabbit antibody against TRF2 (#NB110-57130SS, Novus Biologicals, 1:100 dilution), mouse antibody against RNA pol II (#5408, Abcam, 1:400 dilution), rabbit antibody against RAD51 (#PC130, Calbiochem, 1:50 dilution), rabbit antibody against RPA32/RPA2 (#10359, Abcam, 1:100 dilution), mouse antibody against MLH1 (#51–1327GR, BD PharmigenTM, 1:100 dilution), rabbit antibody against MLH1 (#ab47703, Abcam, 1:100 dilution) and rabbit antibody against γH2AX (#H5912, Sigma-Aldrich, 1:100 dilution).

Techniques: Western Blot

Journal: Journal of Nucleic Acids

Article Title: A ssDNA Aptamer That Blocks the Function of the Anti-FLAG M2 Antibody

doi: 10.4061/2011/720798

Figure Lengend Snippet: 3XABA oligonucleotide blocks the interaction of Flag-tagged proteins with the M2 antibody. (a)-(b) The 3XABA oligo blocks the binding of Flag-TRF2 ΔB to M2-coated beads. Magnetic beads coated with the M2 antibody were incubated in the absence (No Comp) or presence of the indicated competitor (3XABA, 3XCTR, 3XFLAG). In vitro translated [ 35 S]-labeled Flag-TRF2 ΔB was then added and the amount captured by the beads was determined by SDS-PAGE electrophoresis and exposure to a PhophorImager cassette (a). In a second experiment done in triplicate, the amount of [ 35 S]-labeled protein captured was counted by scintillation (b). The amount of [ 35 S]-labeled protein captured in the absence of competitor (No Comp) was arbitrarily set to 100%. In both experiments, beads coated with normal mouse IgG were included as negative control for the capture (IgG). Data represent the mean ± S.D. ( n = 3). (c)-(d) The 3XABA oligo elutes the Flag-TRF2 ΔB proteins already bound to M2-coated beads. The [ 35 S]-Flag-TRF2 ΔB protein was first captured by magnetic beads coated with the M2 antibody. The beads were then incubated in the absence (No comp) or presence of the indicated competitor (3XABA, 3XCTR, 3XFLAG). The amount of [ 35 S]-Flag-TRF2 ΔB released was determined by SDS-PAGE electrophoresis and exposure to a PhophorImager cassette (c). In a second experiment done in triplicate, the amount of [ 35 S]-labeled protein released was counted by scintillation (d). The amount of [ 35 S]-labeled protein released by the boiling (total) was arbitrarily set to 100%. In both experiments, beads boiled to release to all of the captured [ 35 S]-labeled protein were included as positive control for the elution (Total). Data represent the mean ± S.D. ( n = 3).

Article Snippet: Normal mouse IgG (cat. # sc-2025) and anti-vimentin mouse monoclonal antibody (IgG 1 clone sc-6260) were obtained from Santa Cruz (Santa Cruz, CA), as was the rabbit polyclonal antibody against TRF2 (cat. # H-300).

Techniques: Binding Assay, Magnetic Beads, Incubation, In Vitro, Labeling, SDS Page, Electrophoresis, Negative Control, Positive Control

Journal: Oncogene

Article Title: A human cell line that maintains telomeres in the absence of telomerase and of key markers of ALT.

doi: 10.1038/sj.onc.1208934

Figure Lengend Snippet: Figure 5 C3-cl6 cells have lost most markers of classical ALT. (a) C3-cl6 cells do not have APBs. VA13-C3 and C3-cl6 cells were stained for the PML protein (green) and the telomeric protein TRF2 (red) and APBs were detected by the co-localization of the signals (merge). C3-cl6 cells were stained both at early (PD 32) and late (PD 200) PDs. (b) C3-cl6 cells do not have extra-chromosomal telomeric circles. The presence of telomeric circles was analysed by 2D-PFGE and hybridization with a telomeric probe. 25 mg of genomic DNA from GM847 and VA13-C3 cells and 35 mg of genomic DNA from C3-cl6 cells were used. GM847 cells were used as a positive control. The arrows indicate circular telomeric DNA. The hybridization signal in C3-cl6 cells is weaker than in parental and GM847 cells due to the presence of shorter telomeres

Article Snippet: Immunofluorescence For co-localization of PML and TRF2, as previously described (Cerone et al., 2001), cells fixed in 1% formaldehyde and permeabilized with 0.25% Triton X-100 were incubated overnight with a goat polyclonal antibody against PML (N19, Santa Cruz, USA; 2mg/ml) and a rabbit polyclonal antibody against TRF2 (Grobelny et al., 2000) (1 : 50 dilution).

Techniques: Staining, Hybridization, Positive Control