TatC (lack Ser46Phe) and Tat variants (TatN12, TatD60, TatVT6) were cloned into: (a) mammalian expression vector pCMV-myc vector (Clonetech) under the CMV promoter for functional studies, and (b) prokaryotic expression vector pGEX-4T-2 (Invitrogen) to obtain GST-tagged proteins. HIV-1 subtype B TAR was cloned in pcDNA3.1 (Invitrogen) for TAR synthesis to determine Tat–TAR binding activity. Anti-Tat antibody (NIH AIDS Reagent Programme), Anti-myc antibody (Clontech), Anti-GAPDH antibody (Cell Signaling Technology), Anti-rabbit IgG conjugated to HRP (Jackson Immunoresearch), and Anti-Mouse IgG conjugated to HRP (Jackson Immunoresearch) were used in western blotting.
Anti myc antibody
Manufactured by Takara Bio
Sourced in United States
The Anti-Myc antibody is a laboratory reagent used to detect the presence of the Myc protein in biological samples. It is a highly specific and sensitive tool for identifying Myc-tagged proteins in various applications, such as Western blotting, immunoprecipitation, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Pcmv myc vector, by Takara Bio (1 mentions)
Pgex 4t 2, by Thermo Fisher Scientific (1 mentions)
Anti rabbit igg conjugated to hrp, by Jackson ImmunoResearch (1 mentions)
Anti mouse igg conjugated to hrp, by Jackson ImmunoResearch (1 mentions)
Anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
E220 system, by Covaris (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
Supersignal westfemto ecl reagent, by Thermo Fisher Scientific (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Las 3000, by Fujifilm (1 mentions)
4 protocols using anti myc antibody
1
Functional Studies of HIV-1 Tat Variants
2
Chromatin Immunoprecipitation and Quantification
3
Characterization of MORC2 Protein Interactions
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Co-immunoprecipitation was performed with postnatal day 20 mouse testes using affinity-purified MORC2B antibodies as previously described [34 (link)]. Immunoprecipitated proteins were resolved by SDS-PAGE. The protein bands specific to the wild type testis sample were subjected to mass spectrometry for protein identification.
The full-length coding sequences of mouse Morc2a and Morc2b were cloned into pcDNA3.1/myc-His vector and pcDNA3.1/V5-His-TOPO vector respectively. Plasmids were transfected into HEK 293T cells. Forty-eight hours after transfection, cells were collected and lysed in whole cell lysis buffer (50 mM HEPES, pH 7.5, 140 mM NaCl,1 mM DTT, 10% glycerol, 0.5% NP-40, 1 mM PMSF). Immunoprecipitation on protein lysate was performed with anti-V5 antibody (Catalogue number R96025, Invitrogen), followed by Western blotting with anti-Myc antibody (Catalogue number 631206, Clontech).
The full-length coding sequences of mouse Morc2a and Morc2b were cloned into pcDNA3.1/myc-His vector and pcDNA3.1/V5-His-TOPO vector respectively. Plasmids were transfected into HEK 293T cells. Forty-eight hours after transfection, cells were collected and lysed in whole cell lysis buffer (50 mM HEPES, pH 7.5, 140 mM NaCl,1 mM DTT, 10% glycerol, 0.5% NP-40, 1 mM PMSF). Immunoprecipitation on protein lysate was performed with anti-V5 antibody (Catalogue number R96025, Invitrogen), followed by Western blotting with anti-Myc antibody (Catalogue number 631206, Clontech).
4
Egr3 Overexpression and Knockdown in 293T Cells
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293T cells were co-transfected with pcDNA3.1/Myc-Egr3 plasmid and Egr3 siRNA. Cells transfected with empty plasmid or non-targeting siRNA served as negative controls. Cell lysates were prepared in RIPA buffer (50 mM Tris [pH 7.5], 150 mM NaCl, 1% Nonidet P-40, 0.5% Na-deoxycholate, 0.1% SDS, 1 mM dithiothreitol (DTT), 1 mM phenylmethylsulfonyl fluoride (PMSF), 1X Protease Inhibitor [Roche, Indianapolis, IN, USA]) and centrifuged at 13,000 rpm. The supernatant was subjected to bicinchoninic acid (BCA) assays (Thermo Scientific, Rockford, IL, USA) for quantitation. Fifty micrograms of protein were loaded per lane and separated in 10% SDS-polyacrylamide gel electrophoresis (PAGE) gels. Western blotting was performed as previously described [2 (link)]. Anti-α-tubulin (Sigma-Aldrich, USA) and anti-Egr3 (sc-191; Santa Cruz Biotechnology, USA) antibodies were used at 1:2,000 and 1:500, respectively. Anti-Myc antibody (#631206; Clontech, USA) was used at 1:2,000. Horseradish peroxidase (HRP) -conjugated goat anti-rabbit and goat anti-mouse antibodies (GeneDEPOT, Barker, TX, USA) were used at 1:10,000. Super Signal West Femto ECL reagent (Thermo Scientific, USA) was used. Chemiluminescence signal was detected by LAS3000 (FUJIFILM, Tokyo, Japan).
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