S tag

Manufactured by Abcam

The S-tag is a small peptide tag that can be used for the detection and purification of recombinant proteins. It is derived from the S-protein, a ribonuclease inhibitor from bovine pancreas. The S-tag can be genetically fused to the N- or C-terminus of a target protein, allowing for its specific recognition and affinity-based purification.

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Lab products found in correlation

4 protocols using s tag

Western Blot Analysis of HTLV-1 Proteins

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Cells were lysed in NP-40 lysis buffer containing a protease inhibitor cocktail (Millipore Sigma) and quantitated using a Pierce bicinchoninic acid protein assay kit (Thermo Fisher Scientific). Equivalent amounts of protein were separated in Mini-Protean TGX precast 4 to 20% gels (Bio-Rad Laboratories, Hercules, CA, USA) and transferred to nitrocellulose membranes. Membranes were blocked in phosphate-buffered saline (PBS) containing 5% milk plus 0.1% Tween^® 20 and incubated with the primary antibody. The following primary antibodies were used: HBZ (1:1000) [14 (link)], APH-2 (1:1000) [74 (link)], S-tag (1:1000; Abcam), Tax (1:1000), YBX1 (1:5000, Bethyl A303-231A), myc (1:1000; Abcam ab32), and β-actin (1:5000; Millipore Sigma). The secondary antibodies used were horseradish peroxidase-labeled goat anti-rabbit and goat anti-mouse immunoglobulin antibodies (1:5000; Santa Cruz Biotechnology). Membranes were developed using Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) and imaged using an Amersham Imager 600 (GE Healthcare Life Sciences, Piscataway, NJ, USA). Densitometric data were calculated using the ImageQuant TL program (GE Healthcare Life Sciences).

Smith S., Seth J., Midkiff A., Stahl R., Syu Y.C., Shkriabai N., Kvaratskhelia M., Musier-Forsyth K., Jain P., Green P.L, & Panfil A.R. (2023). The Pleiotropic Effects of YBX1 on HTLV-1 Transcription. International Journal of Molecular Sciences, 24(17), 13119.

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Antibody Database for Research

Cited 1 time

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The following primary antibodies were used: S-tag (catalogue number ab184223; Abcam); CBD (E8034S; New England BioLabs); Flag (F1804; Sigma); β-actin (A5316; Sigma); LANA (13-210-100; Advanced Biotechnologies); USP7 (A300-033A [rat]; Bethyl Laboratories); V5-tag (R960-25; Thermo Fisher Scientific); and LDH, HA, His₆, GST, and p53 from Santa Cruz Biotechnologies (catalogue numbers sc-33781, sc-7392, sc-803, sc-138, and sc-126, respectively). vIL-6 rabbit polyclonal antiserum was reported previously (64 (link)). Mouse monoclonal antibody to vIRF-2 (39 (link)) was provided by T. Schulz. Horse radish peroxidase (HRP)-conjugated anti-rabbit IgG and anti-mouse IgG were from Cell Signaling Technology (catalogue numbers 7074S and 7076S, respectively).

Xiang Q., Ju H, & Nicholas J. (2020). USP7-Dependent Regulation of TRAF Activation and Signaling by a Viral Interferon Regulatory Factor Homologue. Journal of Virology, 94(2), e01553-19.

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Western Blot Analysis of PR-Set7 and Riz1 Interaction

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Western analysis was performed using the following antibodies and dilutions: PR-Set7 (Cell Signaling; 1:1K), H4K20 methyl-specific (Active Motif; 1:5K), H3K9 methyl-specific (Millipore; 1:10K), H4 (Abcam; 1:60K), H3 (Abcam; 1:100K), HA (Roche; 1:1K), Myc (Roche; 1:1K), FLAG (Sigma; 1:2.5K), Riz1 [Abcam ab3790 (Figure 1) or ab9710 (Figures 3 and 6); 1:250], His (Novagen; 1:1K), S-tag (Abcam; 1:1K), GST (Millipore; 1:5K), Gal4-DBD (Santa Cruz; 1:5K), G9a (Millipore; 1:1K) and HP1β (Millipore; 1:5K).
Figure 1.

PR-Set7 selectively and directly binds Riz1. (A) Western analysis using indicated antibodies on HeLa whole cell lysates transfected with control or PR-Set7 shRNA (left), or a control or catalytically dead PR-Set7 (CD) plasmid (right). (B) HeLa cells co-transfected with the indicated plasmids were immunoprecipitated (FLAG or HA) followed by Western analysis of bound proteins. (C) HA-immunoprecipitations of HeLa nuclear extracts expressing HA-PR-Set7 or HA-p53 incubated with in vitro translated ³⁵S-Riz1 were fractionated by SDS-PAGE before Western analysis (top) or autoradiography (bottom). (D) S-tag-immunoprecipitations of recombinant His-S-PR-Set7 incubated in vitro with ³⁵S-Riz1 or ³⁵S-G9a. Bound material were fractionated by SDS-PAGE before Western analysis (top) or autoradiography (bottom).

Congdon L.M., Sims J.K., Tuzon C.T, & Rice J.C. (2014). The PR-Set7 binding domain of Riz1 is required for the H4K20me1-H3K9me1 trans-tail ‘histone code’ and Riz1 tumor suppressor function. Nucleic Acids Research, 42(6), 3580-3589.

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Firefly Luciferase Assay for IFN-γ Signaling

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The firefly luciferase reporter plasmid for IFN-γ-responsive promoter and the Renilla luciferase control plasmid (pRL-TK) (31 (link)–33 (link)) were kindly provided by Dr. Hong-Bing Shu (Wuhan University, China). The NSs or NP expression plasmids were constructed as described previously (10 (link), 13 (link), 18 (link)). Mouse anti-NSs antiserum or rabbit antisera to NSs, NP, GP (N-terminal domain), or RdRp were respectively raised against the corresponding viral proteins generated by Escherichia coli (10 (link), 18 (link)). Rabbit antibodies to S-tag (Abcam), STAT1 (Cell Signaling Technology), or STAT2 (Santa Cruz Biotech) and mouse antibodies to HA-tag (Beyotime), β-actin (Beyotime), or STAT1 (Santa Cruz Biotech) were purchased from the indicated manufacturers. Secondary antibodies include goat anti-mouse IgG-fluorescein isothiocyanate (FITC) (Proteintech), goat anti-rabbit IgG-Rhodamine (Chemicon), and goat anti-rabbit IgG-Cy5 (Abcam). Recombinant mouse or human IFN-γ proteins were purchased from Cell Sciences or Peprotech Inc., respectively. Recombinant human IFN-α was from PBL Biomedical Laboratories.

Ning Y.J., Mo Q., Feng K., Min Y.Q., Li M., Hou D., Peng C., Zheng X., Deng F., Hu Z, & Wang H. (2019). Interferon-γ-Directed Inhibition of a Novel High-Pathogenic Phlebovirus and Viral Antagonism of the Antiviral Signaling by Targeting STAT1. Frontiers in Immunology, 10, 1182.

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