rabbit polyclonal anti gamma tubulin  (Abcam)

Journal: BMC Developmental Biology

Article Title: CDC2/SPDY transiently associates with endoplasmic reticulum exit sites during oocyte maturation

doi: 10.1186/1471-213X-9-8

Figure Lengend Snippet: Listing of markers used to establish the identity of cortical structures in immature oocytes.

Article Snippet: The following antibodies and reagents were used (concentration or dilution and catalog number in brackets): mouse monoclonal anti-CDC2 (1 μg/ml; sc-54) and goat polyclonal anti-SEC23 (2 μg/ml; sc-12107) from Santa Cruz Biotechnologies (Santa Cruz, CA, USA); rabbit polyclonal anti-Calnexin (1:250; SPA-860) from Stressgen Biotechnologies (San Diego, CA, USA); mouse monoclonal anti-PSTAIR (1 μg/ml; ab10345, which is directed against EGVPSTAIREISLLKE, a conserved region in cyclin-dependent kinases (CDKs) [ , ]) and rabbit polyclonal anti-gamma-tubulin (1:1000; ab11321) from Abcam (Cambridge, UK); mouse monoclonal anti-NUP153 (4 μg/ml; MMS-102P) from Covance (Berkeley, CA, USA); mouse monoclonal anti-GM130 (1 μg/ml; G65120) and mouse monoclonal anti-cyclin B1 (5 μg/ml; 554179) from BD Biosciences (San Jose, CA, USA); rabbit polyclonal anti-SPDY (2.2 μg/ml; NB100–2521, directed against a conserved region that is present in both isoforms) from Novus Biologicals (Littleton, CO, USA); Mitotracker Deep Red (200 nM; M22426), goat anti-mouse IgG alexa488, goat anti-rabbit IgG alexa568, and rabbit anti-goat IgG alexa488 (20 μg/ml; A11029, A11036, and A11078, respectively) from Molecular Probes (Eugene, OR, USA); donkey anti-mouse IgG Cy3 (6.25 μg/ml; 715-165-151) from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA); rabbit polyclonal anti-P-GM130 (1:100; detects GM130 phosphorylated on serine 25 [ ]) was a generous gift from Dr. Martin Lowe (University of Manchester, UK).

Techniques: Binding Assay

Journal: BMC Developmental Biology

Article Title: CDC2/SPDY transiently associates with endoplasmic reticulum exit sites during oocyte maturation

doi: 10.1186/1471-213X-9-8

Figure Lengend Snippet: Gamma-tubulin, mitochondria, and NUP153 do not associate with ERES . (A-C) 0 h GV stage oocyte labeled for CDC2 (A, green), DNA (A, blue), and gamma-tubulin (B, red). The CDC2-labeled ERES cluster did not label with gamma-tubulin (C). (D-F) 0 h GV stage oocyte labeled for CDC2 (D, green), DNA (D, blue), and mitochondria, using the mitochondrial marker mitotracker (E, red). Mitotracker did not label CDC2-positive ERES (F). (G-I) Confocal section through the same oocyte as in D-F showing the GV. Note that the much lower staining intensity in G-I compared to D-F is solely the result of a ~65 μm change in depth of imaging, since acquisition settings and image enhancement were identical between these images. (J) 0 h GV stage oocyte labeled for NUP153 (green) and DNA (blue). None of the oocytes examined (0/15) showed a cortical domain. The 3 separate double lines around the GV are the result of small size and shape changes between the 3 consecutive sections used to produce the image. Images are either Z-projections of 3 consecutive sections (A-C, J), or single sections (D-I); scale bars represent 20 μm in C, F, I, and J. Arrows indicate CDC2-labeled ERES, and arrowheads denote the position of the GV.

Article Snippet: The following antibodies and reagents were used (concentration or dilution and catalog number in brackets): mouse monoclonal anti-CDC2 (1 μg/ml; sc-54) and goat polyclonal anti-SEC23 (2 μg/ml; sc-12107) from Santa Cruz Biotechnologies (Santa Cruz, CA, USA); rabbit polyclonal anti-Calnexin (1:250; SPA-860) from Stressgen Biotechnologies (San Diego, CA, USA); mouse monoclonal anti-PSTAIR (1 μg/ml; ab10345, which is directed against EGVPSTAIREISLLKE, a conserved region in cyclin-dependent kinases (CDKs) [ , ]) and rabbit polyclonal anti-gamma-tubulin (1:1000; ab11321) from Abcam (Cambridge, UK); mouse monoclonal anti-NUP153 (4 μg/ml; MMS-102P) from Covance (Berkeley, CA, USA); mouse monoclonal anti-GM130 (1 μg/ml; G65120) and mouse monoclonal anti-cyclin B1 (5 μg/ml; 554179) from BD Biosciences (San Jose, CA, USA); rabbit polyclonal anti-SPDY (2.2 μg/ml; NB100–2521, directed against a conserved region that is present in both isoforms) from Novus Biologicals (Littleton, CO, USA); Mitotracker Deep Red (200 nM; M22426), goat anti-mouse IgG alexa488, goat anti-rabbit IgG alexa568, and rabbit anti-goat IgG alexa488 (20 μg/ml; A11029, A11036, and A11078, respectively) from Molecular Probes (Eugene, OR, USA); donkey anti-mouse IgG Cy3 (6.25 μg/ml; 715-165-151) from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA); rabbit polyclonal anti-P-GM130 (1:100; detects GM130 phosphorylated on serine 25 [ ]) was a generous gift from Dr. Martin Lowe (University of Manchester, UK).

Techniques: Labeling, Marker, Staining, Imaging