XFL2 cDNA was cloned into pDisplay vector (Invitrogen) as described previously (Guselnikov et al., 2008 (link)). In addition, the complete coding regions of X. laevis FcRγ.a (AF499689) and FcRγ.b cDNAs (EF431895) were cloned into pAP-Tag5 vector (GenHunter). As a result, these constructions encoded XFL2 receptor with N-terminal HA epitope and FcRγ subunits with c-myc epitope at their C-end. 293T cells were transiently transfected with the plasmids. Transfections were carried out using Unifectin 56 (IBCH, Moscow, Russia) according o the manufacturer’s protocol. After 72 h transfection, the cells were used for immunocytochemistry and flow cytometry. The surface expression of XFL2 was analyzed with FACSCanto II cytometer (BD Bioscience): live cells were stained with mouse monoclonal 12CA5 anti-HA antibody (Abcam) and goat anti-mouse Ig-FITC (BD Bioscience). Intracellular expression of FcRγ subunits was detected using microscope Axioscop 2 plus (Carl Zeiss): transfected cells were smeared on glass slides, fixed with acetone and stained with anti-c-myc 9E10 monoclonal antibodies (Abcam) and goat anti-mouse IgG-Tex-asRed (Molecular Probes).
Anti c myc 9e10 monoclonal antibodies
Manufactured by Abcam
Anti-c-myc 9E10 monoclonal antibodies are laboratory reagents used in research applications. They specifically bind to the c-myc protein, which is a transcription factor involved in cellular processes such as cell growth and proliferation. These antibodies can be used for detection and analysis of the c-myc protein in various experimental techniques.
Automatically generated - may contain errors
Lab products found in correlation
Axioscop 2 plus, by Zeiss (2 mentions)
Mouse monoclonal 12ca5 anti ha antibody, by Abcam (2 mentions)
Fitc goat anti mouse ig, by BD (2 mentions)
Goat anti mouse igg texas red, by Thermo Fisher Scientific (2 mentions)
Pdisplay vector, by Thermo Fisher Scientific (2 mentions)
Facscanto 2 cytometer, by BD (2 mentions)
2 protocols using anti c myc 9e10 monoclonal antibodies
1
Cloning and Expression of XFL2 and FcRγ Subunits
2
Cloning and Expression of XFL2 and FcRγ Subunits
Check if the same lab product or an alternative is used in the 5 most similar protocols
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XFL2 cDNA was cloned into pDisplay vector (Invitrogen) as described previously (Guselnikov et al., 2008 (link)). In addition, the complete coding regions of X. laevis FcRγ.a (AF499689) and FcRγ.b cDNAs (EF431895) were cloned into pAP-Tag5 vector (GenHunter). As a result, these constructions encoded XFL2 receptor with N-terminal HA epitope and FcRγ subunits with c-myc epitope at their C-end. 293T cells were transiently transfected with the plasmids. Transfections were carried out using Unifectin 56 (IBCH, Moscow, Russia) according o the manufacturer’s protocol. After 72 h transfection, the cells were used for immunocytochemistry and flow cytometry. The surface expression of XFL2 was analyzed with FACSCanto II cytometer (BD Bioscience): live cells were stained with mouse monoclonal 12CA5 anti-HA antibody (Abcam) and goat anti-mouse Ig-FITC (BD Bioscience). Intracellular expression of FcRγ subunits was detected using microscope Axioscop 2 plus (Carl Zeiss): transfected cells were smeared on glass slides, fixed with acetone and stained with anti-c-myc 9E10 monoclonal antibodies (Abcam) and goat anti-mouse IgG-Tex-asRed (Molecular Probes).
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