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Rabbit anti γh2avd

Manufactured by Rockland Immunochemicals
Sourced in Japan, United States

Rabbit anti-γH2AvD is a laboratory reagent that can be used to detect the phosphorylated form of the histone variant H2AvD. It is a polyclonal antibody raised in rabbits and specifically recognizes the phosphorylated serine 139 residue of H2AvD. This antibody can be used in various assays, such as Western blotting, immunohistochemistry, and immunofluorescence, to analyze the presence and localization of phosphorylated H2AvD in biological samples.

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4 protocols using rabbit anti γh2avd

1

Immunofluorescence Antibody Staining Protocol

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The following primary antibodies diluted in PBST were used in these experiments: mouse anti-Delta, mouse anti-Arm (Developmental Studies Hybridoma Bank, Iowa City, IA, USA), 1:200; mouse anti-GFP and rabbit anti-GFP (Molecular Probes, Eugene, OR, USA), 1:1000; rat anti-GFP (Nacalai Tesque Inc., Kyoto. Japan), 1:1000; rabbit anti-γH2AvD (Rockland, Gilbertsville, PA, USA) 1:2000; rabbit anti-pS/TQ (Cell Signaling Technologies, Danvers, MA, USA), 1:1000; rabbit anti-phospho-histone H3 (PH3, Millipore, Billerica, MA, USA), 1:1000; mouse anti-γ-tubulin (Sigma-Aldrich), 1:1000; rabbit anti-β-gal (Upstate Biotechnology Inc., Lake Placid, NY, USA), 1:1000; and anti-CCleaved caspase-3 (Cell Signaling Technologies), 1:1000; rabbit anti-pJNK antibody (Cell Signaling Technologies). The following secondary antibodies diluted in PBST were used: goat anti-rabbit FITC (Jackson ImmunoResearch, West Grove, PA, USA), 1:400; goat anti-rabbit Cy3 (Jackson ImmunoResearch), 1:400; goat anti-mouse FITC (Jackson ImmunoResearch), 1:400; goat anti-mouse Cy3 (Jackson ImmunoResearch), 1:400; goat anti-rat FITC (Jackson ImmunoResearch), 1:400, goat anti-rabbit Alexa Fluor® 647 (Jackson ImmunoResearch), 4′,6-diamidino-2-phenylindole (DAPI, Molecular Probes), 1:1000.
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2

Immunostaining of Drosophila Embryos

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The following primary antibodies diluted in PBST were used in these experiments: mouse anti-Dl, mouse anti-Arm (Developmental Studies Hybridoma Bank, Iowa City, IA, USA), 1:200; mouse anti-GFP and rabbit anti-GFP (Molecular Probes, Eugene, OR, USA), 1:1000; rat anti-GFP (Nacalai Tesque Inc., Kyoto. Japan), 1:1000; rabbit anti-γH2AvD (Rockland, Gilbertsville, PA, USA) 1:2000; rabbit anti-phospho-histone H3 (PH3, Millipore, Billerica, MA, USA), 1:1000; mouse anti-γ-tubulin (Sigma–Aldrich), 1:1000; rabbit anti-H3K9me3 (Millipore, Billerica, MA, USA), 1:200; and, mouse anti-HP1 (DSHB, Iowa City, IA, USA), 1:200. The following secondary antibodies diluted in PBST were used: goat anti-rabbit FITC (Jackson ImmunoResearch, West Grove, PA, USA), 1:400; goat anti-rabbit Cy3 (Jackson ImmunoResearch), 1:400; goat anti-mouse FITC (Jackson ImmunoResearch), 1:400; goat anti-mouse Cy3 (Jackson ImmunoResearch), 1:400; goat anti-rat FITC (Jackson ImmunoResearch), 1:400, goat anti-rabbit Alexa Fluor® 647 (Jackson ImmunoResearch), 4′,6-diamidino-2-phenylindole (DAPI, Molecular Probes), 1:1000.
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3

Immunofluorescence Staining Protocols

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The following primary antibodies (diluted in PBST) were used in our experiments: mouse anti-Dl, mouse anti-Arm (Developmental Studies Hybridoma Bank, Iowa City, IA, USA), 1:200; mouse anti-GFP and rabbit anti-GFP (Molecular Probes, Eugene, OR, USA), 1:1000; rabbit anti-γH2AvD (Rockland, Gilbertsville, PA, USA), 1:2000; rabbit anti-pS/TQ (Cell Signaling Technologies, Danvers, MA, USA), 1:1000; rabbit anti-phospho-histone H3 (PH3, Millipore, Billerica, MA, USA), 1:1000; mouse anti-γ-tubulin (Sigma-Aldrich, St. Louis, MO, USA), at 1:1000 dilution. The following secondary antibodies (diluted in PBST) were used: a goat anti-rabbit fluorescein isothiocyanate (FITC) conjugate (Cappel, Solon, OH, USA), 1:400; a goat anti-rabbit Cy3 conjugate (Jackson ImmunoResearch, West Grove, PA, USA), 1:400; a goat anti-mouse FITC conjugate (Jackson ImmunoResearch), 1:400; and a goat anti-mouse Cy3 conjugate (Jackson ImmunoResearch), 1:400. We also used 4′,6-diamidino-2-phenylindole (DAPI, Molecular Probes) for staining, at a 1:1000 dilution.
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4

Immunofluorescence Antibody Panel

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The following primary antibodies diluted in 1× PBST were used in this study: mouse anti-Dl, mouse anti-Pros, mouse anti-HP1 (Developmental Studies Hybridoma Bank, Iowa City, IA, USA), 1:200; mouse anti-GFP and rabbit anti-GFP (Molecular Probes,), 1:1000; rat anti-GFP (Nacalai Tesque Inc., Kyoto, Japan), 1:1000; rabbit anti-phospho-histone H3 (PH3; Millipore, Billerica, MA, USA), 1:1000; anti-Cleaved caspase-3 (Cell Signaling Technologies, Danvers, MA, USA), 1:1000; rabbit anti-γH2AvD (Rockland, Gilbertsville, PA, USA), 1:2000; mouse anti-γ-tubulin (Sigma-Aldrich), 1:1000; and anti-hVDR antibody (Thermo Fisher Scientific, Cleveland, OH, USA), 1:1000. The following secondary antibodies diluted in 1× PBST were used in this study: goat anti-rabbit FITC; goat anti-rabbit Cy3; goat anti-mouse FITC, goat anti-mouse Cy3, goat anti-rat FITC, and goat anti-rabbit Alexa Fluor® 647 (Jackson ImmunoResearch, West Grove, PA, USA), 1:400.
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