anti α2ap neutralizing goat antibody  (R&D Systems)

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of anti-α2AP neutralizing antibodies on adult hippocampal neurogenesis. a Representative images of immunostaining of BrdU and Ki67. Anti-α2AP neutralizing antibodies or control IgG were intracerebroventricularly injected in 12-week-old C57BL/6J mice 2 h after the first intraperitoneal injection of BrdU, and BrdU was administered at 24-h intervals for 7 days. Coronal brain slices were immunostained with antibodies. Arrow heads indicate Ki67 + cells, and arrows indicate examples of BrdU + /Ki67 − cells. Scale bar: 50 μm. b The numbers of BrdU + /Ki67 − cells in the DG were counted in a blinded manner. c Representative images of immunostaining of Dcx. Scale bar: 200 μm. d The numbers of Dcx + cells in the DG were counted in a blinded manner. The values represent the means ± S.E. (control IgG: n = 4, α2AP Ab: n = 5). Statistical significance was evaluated using Student’s t -test. *P < 0.05

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Immunostaining, Control, Injection

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The Effects of anti-α2AP neutralizing antibodies on spatial memory. a Anti-α2AP neutralizing antibodies or control IgG were intracerebroventricularly injected in 12-week-old C57BL/6J mice after the first day of training in the MWM test. On the second day, mice were repeatedly trained, and then the probe tests were performed 30 min, 1 month and 3 months later. b The results of the training sessions. The latency to the target in each trial was measured. The values represent the mean values of 4 trials in each session. c – e The results of the probe tests at 30 min ( c ), 1 month ( d ) and 3 months ( e ) after training. The time in each quadrant was measured. The values represent the mean ± S.E. (control IgG: n = 10, α2AP Ab: n = 9). Statistical significance was evaluated using an ANOVA with an LSD post-hoc test. **P < 0.01, ns non-significant

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Control, Injection

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of the injection of α2AP on adult hippocampal neurogenesis. a Representative images of immunostaining of BrdU and Ki67. α2AP or saline was intracerebroventricularly injected in 12-week-old C57BL/6J mice, 2 h after the first intraperitoneal injection of BrdU, and BrdU was administered at 24-h intervals for 7 days. Coronal brain slices were immunostained with antibodies. Arrow heads indicate Ki67 + cells. Scale bar: 50 μm. b The numbers of BrdU + /Ki67 − cells in the DG were counted in a blinded manner. c Representative images of immunostaining of Dcx. Scale bar: 200 μm. d The numbers of Dcx + cells in the DG were counted in a blinded manner. The values represent the mean ± S.E. (saline: n = 5, α2AP: n = 6). Statistical significance was evaluated using Student’s t -test. *P < 0.05

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Injection, Immunostaining, Saline

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of the injection of α2AP on spatial memory. a α2AP or saline was intracerebroventricularly injected in 12-week-old C57BL/6J mice after the first day of training in the MWM test. On the second day, mice were repeatedly trained, and then the probe tests were performed 30 min and 1 month later. b The results of the training sessions. The latency to the target in each trial was measured, and the values represent the mean values of 4 trials in each session. c and d The results of the probe tests performed 30 min ( c ) and 1 month ( d ) after training. The time in each quadrant was measured. The values represent the mean ± S.E. (saline: n = 8, α2AP: n = 10). Statistical significance was evaluated using an ANOVA with an LSD post-hoc test. **P < 0.01

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Injection, Saline

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: Correlation of α2AP levels in the brain and spatial working memory between young and old mice. a and b The levels of α2AP and plasmin relative to those of GAPDH in the hippocampus and cerebral cortex of young and old C57BL/6J mice were determined by Western blotting (young mice: 12–16 weeks of age, old mice: > 25 months of age). The band intensity was measured using the NIH ImageJ software program and normalized to that of GAPDH. The values were divided by the mean of the normalized intensity of young mice. The bar graphs represent the mean ± SE (arbitrary units: A.U., young mice: n = 8, old mice: n = 9). Statistical significance was evaluated using Student’s t -test. **P < 0.01. c and d The relative intensity of α2AP to GAPDH and the scores of the Y-maze test were analyzed by Pearson’s correlation test. Correlation was evaluated using contribution rate (r 2 ): values of r 2 > 0.16 were considered to indicate a correlation. The values of young mice and old mice were presented with a blank rhombus and filled rhombus, respectively

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Western Blot, Software

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of α2AP deficiency on age-dependent oxidative stress and neuroinflammation. a and b The levels of 13-HPODE-adducted protein relative to those of GAPDH in the hippocampus and cerebral cortex of young and old WT and α2AP −/− mice were determined by Western blotting (young mice: 11 weeks of age, old mice: 60 weeks of age). The relative intensity was measured using the NIH ImageJ software program and normalized to that of GAPDH. The bar graphs represent the mean ± SE (young WT mice: n = 5, young α2AP −/− mice: n = 4, old WT mice: n = 5, old α2AP −/− mice: n = 4). c The levels of IL-6 mRNA in the hippocampus were shown (young mice: n = 7, old mice: n = 5). Statistical significance was evaluated by an ANOVA with an LSD post-hoc test. *P < 0.05., **P < 0.01

Article Snippet: After the first day of MWM training, mice were anesthetized with 1.8–1.9% isoflurane, and then were injected 200 nM of α2AP (Calbiochem, MA, USA) or 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (AF1470, R&D System, MN, USA) as well as saline or normal goat IgG control, respectively (AB-108-C, R&D System) in a total volume of 20 μL into the lateral ventricle (0.5 mm caudally and 1.0 mm laterally to the bregma and 2.0 mm vertically from the skull surface) with a two-step needle (Star needle; Seiseido, Tokyo, Japan) attached to a glass syringe (As one, Osaka, Japan).

Techniques: Western Blot, Software

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of anti-α2AP neutralizing antibodies on adult hippocampal neurogenesis. a Representative images of immunostaining of BrdU and Ki67. Anti-α2AP neutralizing antibodies or control IgG were intracerebroventricularly injected in 12-week-old C57BL/6J mice 2 h after the first intraperitoneal injection of BrdU, and BrdU was administered at 24-h intervals for 7 days. Coronal brain slices were immunostained with antibodies. Arrow heads indicate Ki67 + cells, and arrows indicate examples of BrdU + /Ki67 − cells. Scale bar: 50 μm. b The numbers of BrdU + /Ki67 − cells in the DG were counted in a blinded manner. c Representative images of immunostaining of Dcx. Scale bar: 200 μm. d The numbers of Dcx + cells in the DG were counted in a blinded manner. The values represent the means ± S.E. (control IgG: n = 4, α2AP Ab: n = 5). Statistical significance was evaluated using Student’s t -test. *P < 0.05

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Immunostaining, Injection

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The Effects of anti-α2AP neutralizing antibodies on spatial memory. a Anti-α2AP neutralizing antibodies or control IgG were intracerebroventricularly injected in 12-week-old C57BL/6J mice after the first day of training in the MWM test. On the second day, mice were repeatedly trained, and then the probe tests were performed 30 min, 1 month and 3 months later. b The results of the training sessions. The latency to the target in each trial was measured. The values represent the mean values of 4 trials in each session. c – e The results of the probe tests at 30 min ( c ), 1 month ( d ) and 3 months ( e ) after training. The time in each quadrant was measured. The values represent the mean ± S.E. (control IgG: n = 10, α2AP Ab: n = 9). Statistical significance was evaluated using an ANOVA with an LSD post-hoc test. **P < 0.01, ns non-significant

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Injection

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of the injection of α2AP on adult hippocampal neurogenesis. a Representative images of immunostaining of BrdU and Ki67. α2AP or saline was intracerebroventricularly injected in 12-week-old C57BL/6J mice, 2 h after the first intraperitoneal injection of BrdU, and BrdU was administered at 24-h intervals for 7 days. Coronal brain slices were immunostained with antibodies. Arrow heads indicate Ki67 + cells. Scale bar: 50 μm. b The numbers of BrdU + /Ki67 − cells in the DG were counted in a blinded manner. c Representative images of immunostaining of Dcx. Scale bar: 200 μm. d The numbers of Dcx + cells in the DG were counted in a blinded manner. The values represent the mean ± S.E. (saline: n = 5, α2AP: n = 6). Statistical significance was evaluated using Student’s t -test. *P < 0.05

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Injection, Immunostaining, Saline

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of the injection of α2AP on spatial memory. a α2AP or saline was intracerebroventricularly injected in 12-week-old C57BL/6J mice after the first day of training in the MWM test. On the second day, mice were repeatedly trained, and then the probe tests were performed 30 min and 1 month later. b The results of the training sessions. The latency to the target in each trial was measured, and the values represent the mean values of 4 trials in each session. c and d The results of the probe tests performed 30 min ( c ) and 1 month ( d ) after training. The time in each quadrant was measured. The values represent the mean ± S.E. (saline: n = 8, α2AP: n = 10). Statistical significance was evaluated using an ANOVA with an LSD post-hoc test. **P < 0.01

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Injection, Saline

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: Correlation of α2AP levels in the brain and spatial working memory between young and old mice. a and b The levels of α2AP and plasmin relative to those of GAPDH in the hippocampus and cerebral cortex of young and old C57BL/6J mice were determined by Western blotting (young mice: 12–16 weeks of age, old mice: > 25 months of age). The band intensity was measured using the NIH ImageJ software program and normalized to that of GAPDH. The values were divided by the mean of the normalized intensity of young mice. The bar graphs represent the mean ± SE (arbitrary units: A.U., young mice: n = 8, old mice: n = 9). Statistical significance was evaluated using Student’s t -test. **P < 0.01. c and d The relative intensity of α2AP to GAPDH and the scores of the Y-maze test were analyzed by Pearson’s correlation test. Correlation was evaluated using contribution rate (r 2 ): values of r 2 > 0.16 were considered to indicate a correlation. The values of young mice and old mice were presented with a blank rhombus and filled rhombus, respectively

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Western Blot, Software

Journal: Molecular Brain

Article Title: α2-Antiplasmin as a potential regulator of the spatial memory process and age-related cognitive decline

doi: 10.1186/s13041-020-00677-3

Figure Lengend Snippet: The effects of α2AP deficiency on age-dependent oxidative stress and neuroinflammation. a and b The levels of 13-HPODE-adducted protein relative to those of GAPDH in the hippocampus and cerebral cortex of young and old WT and α2AP −/− mice were determined by Western blotting (young mice: 11 weeks of age, old mice: 60 weeks of age). The relative intensity was measured using the NIH ImageJ software program and normalized to that of GAPDH. The bar graphs represent the mean ± SE (young WT mice: n = 5, young α2AP −/− mice: n = 4, old WT mice: n = 5, old α2AP −/− mice: n = 4). c The levels of IL-6 mRNA in the hippocampus were shown (young mice: n = 7, old mice: n = 5). Statistical significance was evaluated by an ANOVA with an LSD post-hoc test. *P < 0.05., **P < 0.01

Article Snippet: On the next day of the Y-maze test, an injection cannula (AMI-4.5; Eicom) was connected through polyethylene tubing to a Hamilton syringe that had been preloaded with 0.1 μg/μL of an anti-α2AP neutralizing goat antibody (R&D System) or normal goat IgG control (R&D System), and inserted into the guide cannula in the awake mice.

Techniques: Western Blot, Software