anti cstf77 Search Results


mouse monoclonal anti-cstf-77  (Abnova)
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Abnova mouse monoclonal anti-cstf-77
βCstF-64 interacts with <t>CstF-77</t> in PC-12 cells. A) PC-12 cells were seeded onto 10 cm dishes and transfected with 10 μg of 3×FLAG (lanes 1–3), 3×FLAG-CstF-64 (lanes 4–6) or 3×FLAG-βCstF-64 (lanes 7–9) using Lipofectamine 2000. Cells were harvested 48 hours post-transfection. An aliquot of cell lysate (5%) was used as “Total extract” control (upper and lower panels, lanes 1, 4, and 7) while the remaining was added to anti-FLAG antibody coupled to agarose resin and incubated overnight at 4°C. Beads were centrifuged and an aliquot of the supernatant (5%) was used to analyze “unbound” proteins (upper and lower panels, lanes 3, 6, and 9). After extensive washing with NET-2 buffer, “bound” proteins were eluted by boiling in SDS loading buffer. The eluted proteins (50%) were resolved by SDS-PAGE, transferred to nitrocellulose membrane and subjected to immunoblot analysis (IB) using anti-FLAG (upper panel) or anti-CstF-77 antibodies (lower panel). The apparent molecular masses of the 3×FLAG-CstF-64 and βCstF-64 proteins and CstF-77 are indicated on left. B, C. Exogenously expressed CstF-64 and βCstF-64 proteins localize to the nucleus. PC-12 cells were grown on coverslips in 24-well plates and transfected with FLAG-MS2-CstF-64 (panel A) or FLAG-MS2-βCstF-64 using Lipofectamine 2000. After 48 hours, cells were fixed with paraformaldehyde, permeabilized with Triton-X-100 and incubated with anti-FLAG antibody for two hours. Secondary antibody conjugated to Alexa fluor 488 (green) was added and slides containing these coverslips were subsequently used for photomicrography. The images shown above are an overlay of the phase contrast (red) with the fluorescent signal emanating from the FLAG-CstF-64 and βCstF-64 proteins. Arrows indicate anti-FLAG staining of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C), predominantly in nuclei; arrowheads denote less intense cytoplasmic staining, indicating a paucity of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C) in cytoplasm.
Mouse Monoclonal Anti Cstf 77, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+cstf77/mouse+monoclonal+anti+cstf+77/pmc03783208-77-30-33
Average 90 stars, based on 1 article reviews
mouse monoclonal anti-cstf-77 - by Bioz Stars, 2026-09
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βCstF-64 interacts with CstF-77 in PC-12 cells. A) PC-12 cells were seeded onto 10 cm dishes and transfected with 10 μg of 3×FLAG (lanes 1–3), 3×FLAG-CstF-64 (lanes 4–6) or 3×FLAG-βCstF-64 (lanes 7–9) using Lipofectamine 2000. Cells were harvested 48 hours post-transfection. An aliquot of cell lysate (5%) was used as “Total extract” control (upper and lower panels, lanes 1, 4, and 7) while the remaining was added to anti-FLAG antibody coupled to agarose resin and incubated overnight at 4°C. Beads were centrifuged and an aliquot of the supernatant (5%) was used to analyze “unbound” proteins (upper and lower panels, lanes 3, 6, and 9). After extensive washing with NET-2 buffer, “bound” proteins were eluted by boiling in SDS loading buffer. The eluted proteins (50%) were resolved by SDS-PAGE, transferred to nitrocellulose membrane and subjected to immunoblot analysis (IB) using anti-FLAG (upper panel) or anti-CstF-77 antibodies (lower panel). The apparent molecular masses of the 3×FLAG-CstF-64 and βCstF-64 proteins and CstF-77 are indicated on left. B, C. Exogenously expressed CstF-64 and βCstF-64 proteins localize to the nucleus. PC-12 cells were grown on coverslips in 24-well plates and transfected with FLAG-MS2-CstF-64 (panel A) or FLAG-MS2-βCstF-64 using Lipofectamine 2000. After 48 hours, cells were fixed with paraformaldehyde, permeabilized with Triton-X-100 and incubated with anti-FLAG antibody for two hours. Secondary antibody conjugated to Alexa fluor 488 (green) was added and slides containing these coverslips were subsequently used for photomicrography. The images shown above are an overlay of the phase contrast (red) with the fluorescent signal emanating from the FLAG-CstF-64 and βCstF-64 proteins. Arrows indicate anti-FLAG staining of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C), predominantly in nuclei; arrowheads denote less intense cytoplasmic staining, indicating a paucity of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C) in cytoplasm.

Journal: Gene

Article Title: Polyadenylation Site-specific Differences in the Activity of the Neuronal βCstF-64 Protein in PC-12 Cells

doi: 10.1016/j.gene.2013.08.007

Figure Lengend Snippet: βCstF-64 interacts with CstF-77 in PC-12 cells. A) PC-12 cells were seeded onto 10 cm dishes and transfected with 10 μg of 3×FLAG (lanes 1–3), 3×FLAG-CstF-64 (lanes 4–6) or 3×FLAG-βCstF-64 (lanes 7–9) using Lipofectamine 2000. Cells were harvested 48 hours post-transfection. An aliquot of cell lysate (5%) was used as “Total extract” control (upper and lower panels, lanes 1, 4, and 7) while the remaining was added to anti-FLAG antibody coupled to agarose resin and incubated overnight at 4°C. Beads were centrifuged and an aliquot of the supernatant (5%) was used to analyze “unbound” proteins (upper and lower panels, lanes 3, 6, and 9). After extensive washing with NET-2 buffer, “bound” proteins were eluted by boiling in SDS loading buffer. The eluted proteins (50%) were resolved by SDS-PAGE, transferred to nitrocellulose membrane and subjected to immunoblot analysis (IB) using anti-FLAG (upper panel) or anti-CstF-77 antibodies (lower panel). The apparent molecular masses of the 3×FLAG-CstF-64 and βCstF-64 proteins and CstF-77 are indicated on left. B, C. Exogenously expressed CstF-64 and βCstF-64 proteins localize to the nucleus. PC-12 cells were grown on coverslips in 24-well plates and transfected with FLAG-MS2-CstF-64 (panel A) or FLAG-MS2-βCstF-64 using Lipofectamine 2000. After 48 hours, cells were fixed with paraformaldehyde, permeabilized with Triton-X-100 and incubated with anti-FLAG antibody for two hours. Secondary antibody conjugated to Alexa fluor 488 (green) was added and slides containing these coverslips were subsequently used for photomicrography. The images shown above are an overlay of the phase contrast (red) with the fluorescent signal emanating from the FLAG-CstF-64 and βCstF-64 proteins. Arrows indicate anti-FLAG staining of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C), predominantly in nuclei; arrowheads denote less intense cytoplasmic staining, indicating a paucity of FLAG-CstF-64 (panel B) or FLAG-βCstF-64 (panel C) in cytoplasm.

Article Snippet: 2.2 Antibodies The following antibodies were used in this study: anti-α-tubulin and anti-FLAG antibodies (Sigma, St. Louis, MO); mouse monoclonal anti-CstF-64 (3A7, [ 17 ]); rabbit anti-βCstF-64 [ 15 ]; mouse monoclonal anti-CstF-77 (Abnova, Walnut, CA, catalog #H00001479-M01); and rabbit polyclonal anti-CstF-77 (Bethyl, Montgomery, TX, catalog #A301-096A).

Techniques: Transfection, Control, Incubation, SDS Page, Membrane, Western Blot, Staining