rabbit polyclonal antibody against cdk5  (Santa Cruz Biotechnology)

Journal: The Journal of Neuroscience

Article Title: Loss of Cyclin-Dependent Kinase 5 from Parvalbumin Interneurons Leads to Hyperinhibition, Decreased Anxiety, and Memory Impairment

doi: 10.1523/JNEUROSCI.0969-14.2015

Figure Lengend Snippet: Genetic ablation of Cdk5 specifically in PV interneurons. A, B, Immunoreactivity for PV and Cdk5 in the motor cortex in 3.5-month-old fCdk5 (control) and PVCre;fCdk5 (mutant) littermate mice. The expression of Cdk5 (green) is present in all PV-positive (red) cells (arrows) in control mice (A). The complete ablation of Cdk5 is observed in PV interneurons of the PVCre;fCdk5 mice (B). Scale bars, 15 μm. C, D, Immunoreactivity for PV interneurons in different brain areas and layers in the cortex in control and PVCre;fCdk5 mice. C, PV interneurons (red) in different layers of the motor cortex in control and PVCre;fCdk5 mice, with Cux I (green) immunoreactivity labeling cortical layers II–IV. Scale bars, 100 μm. D, Bar graphs represent the relative numbers of PV interneurons in the cingulate cortex, hippocampus, and different layers of the motor cortex in 3.5-month-old control and PVCre;fCdk5 littermate mice. E, Representative Western blot and related quantification of Cdk5 protein in control and PVCre;fCdk5 (red) brains. Black, Control mice; red, PVCre;fCdk5 mice; ns, not significant. p > 0.05, error bars indicate ±SEM.

Article Snippet: The following primary antibodies were used: PV-positive interneurons were identified with anti-PV rabbit polyclonal PV-25 antibody at 1:2000 [catalog #PV 25; Resource Research Identifier (RRID) AB10000344; Swant] and mouse monoclonal PV-235 antibody at 1:1000 (catalog #PV 235; RRID AB10000343; Swant); rabbit polyclonal antibody against Cux I at 1:200 (M 22; catalog #sc-13024; RRID AB2261231; Santa Cruz Biotechnology) was used as a cortical layer marker; and Cdk5 immunoreactivity was identified with C8 rabbit polyclonal antibody against Cdk5 at 1:100 (catalog #sc-173; RRID AB631224 , Santa Cruz Biotechnology).

Techniques: Control, Mutagenesis, Expressing, Labeling, Western Blot

Journal: British Journal of Pharmacology

Article Title: Neuroprotective effects of the anti‐cancer drug sunitinib in models of HIV neurotoxicity suggests potential for the treatment of neurodegenerative disorders

doi: 10.1111/bph.12875

Figure Lengend Snippet: Table 1

Article Snippet: Membranes were incubated with rabbit polyclonal antibodies against CDK5 (1:500, C-8, Santa Cruz Biotechnology, Santa Cruz, CA, USA), phosphorylated CDK5 (Tyr 15 , 1:500, Santa Cruz Biotechnology) and p35/p25 (1:500, C-19, Santa Cruz Biotechnology).

Techniques: Activity Assay

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Immunoblot analysis of levels of CDKs and phosphorylated tau (pTau) expression in HIV encephalopathy (HIVE) cases. Western blot analysis was performed with frontal cortex homogenates from HIV-positive cases without encephalitis and from cases with HIVE. For each lane, 15 μg of the membrane fraction was used. A: Representative blot probed with antibodies against CDK5, pCDK5, p35/p25, CDK1, cyclin-D, and CDK2. B and C: Analysis of the immunoreactivity levels of CDKs and activators, expressed as ratios relative to actin levels (to correct for loading). D: Representative blot probed with antibodies against total tau (tTau), pTau Ser396 and Ser404 (PHF-1), pTau Ser202 (AT8), and pTau Thr231 (CP9). E: Analysis of the levels of tau immunoreactivity, expressed as ratios relative to actin levels (to correct for loading). *P < 0.01 by unpaired two-tailed Student's t-test (n = 8 cases per group).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Western Blot, Expressing, Membrane, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Immunocytochemical analysis of CDKs and pTau immunoreactivity in HIVE cases. Representative bright-field images are shown from immunolabeled 40-μm-thick microtome sections from the frontal cortex. A and B: CDK5 immunoreactivity in pyramidal neurons in layer III in HIV-positive (A) and HIVE (B) cases. C: Image analysis shows increased corrected optical density values of CDK5 immunoreactivity in HIVE cases, compared with HIV-positive controls. D and E: p35/p25 immunoreactivity in pyramidal neurons in layer III in HIV-positive (D) and HIVE (E) cases. F: Image analysis shows increased corrected optical density values of p35/p25 immunoreactivity in HIVE cases, compared with HIV-positive controls. G and H: CDK1 immunoreactivity in pyramidal neurons in layer III in HIV-positive (G) and HIVE (H) cases. I: Image analysis shows similar CDK1 levels in HIVE and HIV-positive controls. J: PHF-1 immunoreactivity in a control HIV-positive case. K: pTau (PHF-1 antibody) in pyramidal neurons and axons (arrows) in an HIVE case. L: Image analysis shows increased corrected optical density values of pTau (PHF-1) immunoreactivity in HIVE cases, compared with HIV-positive controls. *P < 0.01 by unpaired two-tailed Student's t-test (n = 8 cases per group). Scale bar = 15 μm (all images).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Immunolabeling, Control, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Immunoblot analysis of levels of CDKs and pTau expression in gp120 Tg mice. Western blot analysis was performed with total homogenates from the cortex of non-Tg and gp120 Tg mice. For each lane, 15 μg of the membrane fraction was used. A: Representative blot probed with antibodies against CDK5, pCDK5, p35/p25, CDK1, cyclin-D, and CDK2. B and C: Analysis of the immunoreactivity levels of CDKs and activators, expressed as ratios relative to actin levels o correct for loading. D: Representative blot probed with antibodies against tTau, pTau Ser396 and Ser404 (PHF-1), and pTau Ser202 (AT8). E: Analysis of the levels of tau immunoreactivity, expressed as ratios relative to actin levels (to correct for loading). *P < 0.01 by unpaired two-tailed Student's t-test (n = 8 mice per group, 12 months old).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Western Blot, Expressing, Membrane, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Immunocytochemical analysis of CDKs and pTau immunoreactivity in gp120 Tg mice. All micrographs show representative bright-field images from the frontal cortex. A and B: CDK5 immunoreactivity in pyramidal neurons in layer III in non-Tg (A) and gp120 Tg (B) mice. C: Image analysis shows increased CDK5 immunoreactivity, expressed as corrected optical density, in gp120 Tg mice, compared with non-Tg controls. D and E: p35/p25 immunoreactivity in pyramidal neurons in non-Tg (D) and gp120 Tg (E) mice. F: Image analysis shows increased p35/p25 immunoreactivity, expressed as corrected optical density, in gp120 Tg, compared with non-Tg control mice. G and H: CDK1 immunoreactivity in pyramidal neurons in non-Tg (G) and gp120 Tg (H) mice. I: Image analysis shows similar CDK1 levels in non-Tg and gp120 Tg mice. J: PHF-1 immunoreactivity in non-Tg control mice. K: pTau (PHF-1 antibody) immunoreactivity in pyramidal neurons of gp120 Tg mice. L: Image analysis shows increased pTau (PHF-1) immunoreactivity, expressed as corrected optical density, in gp120 Tg mice, compared with non-Tg controls. *P < 0.01 by unpaired two-tailed Student's t-test (n = 8 mice per group, 12 months old). Scale bar = 15 μm (all images).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Control, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Double-labeled immunocytochemical analysis of CDK5 and pTau colocalization in gp120 Tg mice. Laser scanning confocal micrographs are from the frontal cortex of non-Tg and gp120 Tg mice (12 months old) double-labeled with antibodies against CDK5 (red) and pTau (green). Cell nuclei (blue) were costained with DAPI. A–C: Low levels of CDK5 and pTau (AT8) immunoreactivity in non-Tg mice. D–F: High levels of CDK5 expression and colocalization (arrows) with pTau (AT8) in gp120 Tg mice. G–I: Low levels of CDK5 and pTau (PHF-1) immunoreactivity in non-Tg mice. J–L: High levels of CDK5 expression and colocalization (arrows) with pTau (PHF-1) in gp120 Tg mice. M and N: Semiquantitative analysis of the percentage of cortical neurons displaying colocalization of CDK5 and pTau immunoreactivity with the AT8 (M) or PHF-1 (N) antibodies in cortical neurons from non-Tg and gp120 Tg mice. *P < 0.01 by unpaired two-tailed Student's t-test (n = 8 mice per group, 12 months old). Scale bar = 10 μm (all images).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Labeling, Expressing, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Immunoblot analysis of CDKs and pTau levels in crosses between gp120 Tg and Cdk5+/− mice. Western blot analysis was performed with homogenates from the cortex of animals generated from the crosses. For each lane, 15 μg of the membrane fraction was used. A: Representative blot probed with antibodies against CDK5, pCDK5, p35/p25, CDK1, and cyclin-D. B and C: Analysis of the immunoreactivity levels of CDKs and activators, expressed as ratios relative to actin levels (to correct for loading). D: Representative blot probed with antibodies against tTau, pTau Ser396 and Ser404 (PHF-1), pTau Ser202 (AT8), and pTau Thr231 (CP9). E: Analysis of the levels of tau immunoreactivity, expressed as ratios relative to actin levels (to correct for loading). *P < 0.05 by one-way analysis of variance with post hoc Dunnett's test, compared with control non-Tg mice (n = 8 mice per group, 12 months old). **P < 0.05 by one-way analysis of variance with post hoc Tukey-Kramer test, compared with gp120 Tg mice.

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Western Blot, Generated, Membrane, Control

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Effects of CDK5 knockdown on pTau immunoreactivity in gp120 Tg mice. Micrographs are representative bright-field images from brain sections of non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− mice. A–D: Low-magnification view of the neocortex and hippocampus in brain sections from non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− mice immunostained with an antibody against pTau (PHF-1). E–H: Higher-magnification view shows pyramidal neurons in the neocortex of non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− immunostained with an antibody against pTau (PHF-1). I: Image analysis shows increased levels of pTau (PHF-1) immunoreactivity in gp120 Tg mice, compared with non-Tg and Cdk5+/− mice. J–M: Higher-magnification view of CDK5 immunoreactivity in pyramidal neurons in the neocortex of non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− mice. N: Image analysis shows reduced CDK5 levels, expressed as corrected optical density values, in Cdk5+/− and gp120 Tg/Cdk5+/− mice, compared with non-Tg controls. *P < 0.05 by one-way analysis of variance with post hoc Dunnett's test, compared with control non-Tg mice (n = 8 mice per group, 12 months old). **P < 0.05 by one-way analysis of variance with post hoc Tukey-Kramer test, compared with gp120 Tg. Scale bar: 375 μm (A–D); 25 μm (E–H and J–M).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Knockdown, Control

Journal: The American Journal of Pathology

Article Title: Increased CDK5 Expression in HIV Encephalitis Contributes to Neurodegeneration via Tau Phosphorylation and Is Reversed with Roscovitine

doi: 10.1016/j.ajpath.2010.12.033

Figure Lengend Snippet: Effects of CDK5 knockdown on water maze performance and markers of neurodegeneration in gp120 Tg mice. A: Performance in the visible (days 1 to 3) and hidden (days 4 to 7) portions of the water maze test in crosses between gp120 Tg and Cdk5+/− mice. During the visible portion of the test, mice from all four groups performed with similar abilities; during the hidden part of the test (submerged platform), gp120 Tg mice took a longer path to complete the test, compared with non-Tg mice. The Cdk5+/− and gp120 Tg/Cdk5+/− mice performed similarly to the non-Tg controls. B–E: Laser scanning confocal micrographs show MAP2 immunolabeling of the neuropil in the neocortex of non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− mice. F: Image analysis shows decreased levels of MAP2 immunoreactivity in gp120 Tg mice, compared with non-Tg controls, and recovery in the gp120 Tg/Cdk5+/− mice. G–J: Bright-field micrographs show astroglial cells immunolabeled with antibody against GFAP in the neocortex of non-Tg, gp120 Tg, Cdk5+/−, and gp120 Tg/Cdk5+/− mice. K: Image analysis shows increased GFAP immunoreactivity in gp120 Tg mice, compared with non-Tg, and recovery in the gp120 Tg/Cdk5+/− mice. *P < 0.05 by one-way analysis of variance with post hoc Dunnett's test, compared with control non-Tg mice (n = 8 mice per group, 12 months old). **P < 0.05 by one-way analysis of variance with post hoc Tukey-Kramer test, compared with gp120 Tg. Scale bar = 25 μm (all images).

Article Snippet: Membranes were incubated overnight at 4°C with rabbit polyclonal antibodies against CDK5 (C-8, 1:500), phosphorylated CDK5 (Tyr15, 1:500), p35/p25 (C-19, 1:500), CDK1/2 (AN21.2, 1:1000), or cyclin-D (H-295, 1:1000), all from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Knockdown, Immunolabeling, Control