BirA*-dCas9-EGFP construct was generated by sub-cloning BirA*-HA from pcDNA3.1 MCS-BirA (R118G)-HA (Addgene #36047) into pSLQ1658-dCas9-EGFP (Addgene #51023), BglII and NCOI were selected restriction enzyme digestion sites. SgRNA-Telo construct was created by subcloning telomere repeat sequence into gRNA_Cloning Vector (Addgene #41824) with Gibson Assembly assay. To construct the DSP∆NLS, N-DSP, and C-DSP expression vectors, gene fragments were generated from 1136-Desmoplakin-GFP (Addgene #32227) by PCR, then the PCR fragments were cloned into pLJM1-EGFP. All the antibodies used in this research were as follow: anti-GFP (MBL, M048-3), anti-TRF1 (Santa cruz, sc-56807), anti-POT1 (Santa cruz, sc-81711), anti-TRF2 (Novus Biologicals, NB110-57130), anti-γH2AX(EMD Millipore, 05-636), anti-DSP (Abcam, 118804), anti-Phospho-Chk1 (Ser345) (Cell Signaling technology, #2341). Telo-sgRNA sequence: AGGGTTAGGGTTAGGGTTA.
Anti dsp
Manufactured by Abcam
Anti-DSP is a reagent used for the detection and quantification of DSP (Desmoplakin) protein in various biological samples. It is designed for research use only and its core function is to provide a tool for researchers to study the expression and localization of DSP in their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho chk1 ser345, by Cell Signaling Technology (1 mentions)
Grna cloning vector, by Addgene (1 mentions)
Anti γh2ax, by Merck Group (1 mentions)
Anti trf1, by Santa Cruz Biotechnology (1 mentions)
Anti pot1, by Santa Cruz Biotechnology (1 mentions)
Anti trf2, by Novus Biologicals (1 mentions)
Pslq1658 dcas9 egfp, by Addgene (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal laser scanning microscope, by Olympus (1 mentions)
2 protocols using anti dsp
1
Generating Fluorescent Chromatin Constructs
2
Immunofluorescence Staining of Cellular Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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OLCs in 35 mm glass bottom dishes were fixed with 4% paraformaldehyde for 30 min at room temperature, permeabilized in 0.1% Triton X-100 for 5 min, and blocked with 2% goat serum (cat. no. C0265; Beyotime Biotechnology, Shanghai, China) at 37°C for 1 h. Cells were then incubated with rabbit anti-Hey1 (cat. no. ab22614; 1:50; Abcam) or anti-DSP (cat. no. sc-33587; 1:50; Santa Cruz Biotechnology, Inc.) primary antibody at 4°C overnight. Finally, cells were incubated with Alexa Fluor 594-conjugated goat anti-rabbit IgG secondary antibody (cat. no. A11037; 1:400; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 1 h, and nuclei were stained with 2 μg/ml DAPI (cat. no. C1002; Beyotime Biotechnology) for 5 min at room temperature. Fluorescence was examined using a FV1000 confocal laser scanning microscope (Olympus Corporation, Tokyo, Japan). Fluorescence intensity was determined with Image-Pro Plus 6.0 software (Media Cybernetics, Inc.).
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