Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 1. Cyclin-dependent kinase CDK1/cyclin B kinase activity in prophase and metaphase oocytes from Arenicola marina and Areni- cola defodiens. The kinase was purified from 50 µL packed oo- cytes by affinity chromatography on p9CKShs1-Sepharose beads and assayed in triplicate, using histone H1 as a substrate. Bars, Standard Error (SE).

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Activity Assay, Purification, Affinity Chromatography

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 2. Gel filtration of CDK1 and cyclin B purified from A. marina metaphase I oocyte extracts. The extract was loaded on a Sephacryl S-200 column, and fractions were purified on p9CKShs1-Sepharose beads prior to kinase activity determination (upper section) or cyclin B (middle section) and CDK1 (lower section) detection by Western blotting. The column was calibrated with molecular weight standards.

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Filtration, Purification, Activity Assay, Western Blot, Molecular Weight

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 3. Gel filtration of CDK1 and cyclin B purified from A. marina prophase I oocyte extracts. The extract was loaded on a Sephacryl S-200 column, and fractions were purified on p9CKShs1-Sepharose beads prior to cyclin B (upper panel) and CDK1 (lower panel) detection by Western blotting. Only the representative early, high molecular weight fractions (High MW fractions) and late, low molecular weight fractions (Low MW fractions) are shown.

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Filtration, Purification, Western Blot, High Molecular Weight, Molecular Weight

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 4. Comparison of histone H1 kinase activity of CDK1/ cyclin B and of monomeric CDK1 purified from either prophase or metaphase A. marina oocytes. Extracts were purified by gel filtration on a Sephacryl S-200 column, and fractions were purified on p9CKShs1-Sepharose beads prior to kinase activity determination. Only the activities of the representative early, high molecular weight fractions (High MW fractions), containing CDK1/cyclin B, and the late, low molecular weight fractions (Low MW fractions), containing monomeric CDK1 are shown

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Comparison, Activity Assay, Purification, Filtration, High Molecular Weight, Molecular Weight

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 6. Western blotting of CDK1 affinity-purified on p9CKShs1- sepharose beads from prophase (P) and metaphase (M) oocytes of A. marina and A. defodiens. The p9CKShs1-bound CDK1 was analy- zed by Western blotting using anti-PSTAIR antibodies (A), anti- phospho-cdc2 antibodies (B), anti-phosphotyrosine antibodies (C), or anti-GEGTYG antibodies (D). Note that CDK1 migrates under two forms, the upper (U) and lower (L) bands.

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Western Blot, Affinity Purification

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 7. Roscovitine and olomoucine inhibit CDK1/cyclin B and oocyte maturation in a dose-dependent manner. (A) GVBD in maturing A. marina oocytes treated with increasing concen- trations of each drug. (B) Kinase activity of CDK1/cyclin B affinity-purified from A. marina metaphase oocytes and assayed in the presence of concentrations of each drug. , olomoucine; , roscovitine.

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Activity Assay, Affinity Purification

Journal: Development, growth & differentiation

Article Title: CDK1/cyclin B regulation during oocyte maturation in two closely related lugworm species, Arenicola marina and Arenicola defodiens.

doi: 10.1111/j.1440-169x.2004.00723.x

Figure Lengend Snippet: Fig. 8. Schematic representation of CDK1/cyclin B regulation and activation in Arenicola sp. In prophase oocytes (left), CDK1 is either monomeric or in association with cyclin B. Both forms of the kinase are inactive. Cyclin B– associated CDK1 is maintained inactive by phosphorylation on Tyr15, possibly also on Thr14. In metaphase oocytes (right), only the CDK1/cyclin B heterodimer is fully active. This activation of the kinase comes from the dephos- phorylation of Tyr15, possibly also of Thr14.

Article Snippet: Polyclonal antiphospho-cdc2 (Thr161) antibodies were raised against a synthetic phospho-Thr161 peptide corresponding to residues surrounding Thr161 of human cdc2 in rabbit (Cell Signaling no. 9114).

Techniques: Activation Assay, Phospho-proteomics

Journal: British Journal of Cancer

Article Title: Curcumin–cyclodextrin complexes potentiate gemcitabine effects in an orthotopic mouse model of lung cancer

doi: 10.1038/bjc.2012.379

Figure Lengend Snippet: Curcumin–CD complexes affect the production of proteins related to apoptosis and cell cycle. ( A ) Western blot analyses have been performed to measure cleaved PARP, phospho-cyclin B1 and phospho-cdc2 levels. Actin is shown as loading control. ( B – D ) Quantification was performed by densitometric scanning. Results are expressed as mean±s.e.m. and are representative of two independent experiments.

Article Snippet: After blocking in phosphate-buffered saline containing 10% non-fat milk and Tween 20 (0.05%), membranes were incubated with the primary antibody: anti-poly (ADP-ribose) polymerase (PARP) rabbit monoclonal antibody, anti-cyclin B1 mouse monoclonal antibody, antiphospho-cyclin B1 rabbit polyclonal antibody and antiphospho-cdc2 (Thr161) rabbit polyclonal antibody (Cell Signaling, Leiden, The Netherlands).

Techniques: Western Blot, Control