First, unmodified or K119-malonylated Hta1-C-tail peptides conjugated with biotin were captured by Dynabeads M-280 streptavidin (invitrogen). After washing by PBS three times, the peptide-bound beads were mixed with recombinant GST-Bub1C protein in buffer-P (20 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1% Triton X-100, 1 mg/ml BSA) and incubated at 4 °C for 1 hour. After washing with buffer-P without BSA twice, the bead-bound proteins were analyzed by immunoblotting with an anti-GST antibody (wako, 013-21851).
Anti gst antibody
Manufactured by Fujifilm
Sourced in Japan
The Anti-GST antibody is a lab equipment product developed by Fujifilm. It is a specific antibody that binds to the Glutathione S-Transferase (GST) protein, which is commonly used as a tag in recombinant protein expression and purification experiments.
Automatically generated - may contain errors
Lab products found in correlation
Dynabeads m 280 streptavidin, by Thermo Fisher Scientific (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Nacalai Tesque (1 mentions)
Anti py20, by Santa Cruz Biotechnology (1 mentions)
E coli bl21 de3, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Alkaline phosphatase, by Merck Group (1 mentions)
Pgex 6p 1, by Cytiva (1 mentions)
Phos tag acrylamide aal 107, by Fujifilm (1 mentions)
In fusion hd cloning kit, by Takara Bio (1 mentions)
Gst accept resin, by Nacalai Tesque (1 mentions)
2 protocols using anti gst antibody
1
Bub1C Binding to Malonylated Histone
2
Recombinant Human ABL1 Protein Production
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The authors purchased pGEX-6p-1 from Cytiva (Sheffield, UK). The authors obtained pET21a(+), pCDF-1b and E. coli BL21(DE3) from Merck Millipore (Darmstadt, Germany). Kazusa DNA clone encoding human ABL1 (pF1KB4477) was commercially sourced from Promega (WI, USA). The In-Fusion HD cloning kit was purchased from Takara Bio Inc. (Kusatsu, Japan). Luria-Bertani (LB) medium, IPTG and GST-accept resin were purchased from Nacalai Tesque (Kyoto, Japan). Ampicillin, streptomycin and alkaline phosphatase were obtained from Sigma-Aldrich (MO, USA). Anti-6His antibody, anti-GST antibody and Phos-tag acrylamide AAL-107 were commercially sourced from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). Anti-PY20 was purchased from Santa Cruz Biotechnology (TX, USA). Human PTP1B was obtained from Abcam (Cambridge, UK).
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