sap97 mouse k64 15 igg1  (NeuroMab)

Journal: Cells

Article Title: Age-Dependent Dysregulation of APP in Neuronal and Skin Cells from Fragile X Individuals

doi: 10.3390/cells12050758

Figure Lengend Snippet: Modulation of ADAM10 in human TDI and FXS fibroblasts. ( A ) Schematic representation of the mechanism of action of the TAT-Pro peptide (image created in BioRender.com). ( B ) Left, representative Western blot showing SAP97 expression in TDI and FXS fibroblasts. Right, the bar plot shows the quantification of SAP97 levels in TDI and FXS fibroblasts (TDI n = 8; FXS n = 8). Error bars represent the SEM (** p < 0.01, Mann–Whitney test). ( C ) Left, representative Western blots showing the levels of sAPPα in non-treated fibroblasts (NT), treated with the control peptide (Ala) or with the specific peptide (Pro). Right, the bar plots show the quantification of sAPPα normalized to Coomassie staining (TDI n = 6, FXS n = 10). Error bars represent the SEM (Two-way ANOVA analysis: genotype effect F (1, 42) = 61.97 *** p < 0.001; treatment effect F(2, 42) = 2.866 p > 0.05; interaction effect F(2, 42) = 4.422) (*** p < 0.001 NT TDI vs. NT FXS; *** p < 0.001 TAT-Ala TDI vs. TAT-Ala FXS, Sidak’s multiple comparisons test). ( D ) Left, representative Western blot showing the levels of puromycin incorporation in fibroblasts NT, treated with the Ala or Pro peptides. Right, the bar plots show the quantification of puromycin normalized to Coomassie staining in TDI and FXS fibroblasts expressing regular or high levels of protein synthesis (TDI n = 6, FXS high n = 5; FXS regular n = 5). Error bars represent the SEM (Two-way ANOVA analysis: genotype effect F(2, 39) = 34.87 *** p < 0.001; treatment effect F(2, 39) = 0.4527 p > 0.05; interaction effect F(4, 39) = 2.008 p > 0.05) (*** p < 0.001 NT TDI vs. NT FXS; ** p < 0.01 TAT-Ala TDI vs. TAT-Ala FXS, Sidak’s multiple comparisons test).

Article Snippet: Membranes were incubated using the following specific antibodies, including mouse anti-puromycin (1:500, DSHB), mouse anti-Vinculin (1:2000, Merck), mouse anti-GAPDH (1:2000, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-APP (1:2000, Merck), rabbit anti-ADAM10 (1:500, Abcam, Cambridge, UK), mouse anti-sAPPα (1:500, IBL America, Minneapolis, MN, USA), rabbit anti-OCT3/4 (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-MAP2 (1:2000, Merck), mouse anti-Nestin (1:1000 Santa Cruz Biotechnology), mouse anti-SAP97 (1:1000, ENZO Life Sciences, Farmingdale, NY, USA) and rabbit anti-FMRP (1:1000, produced in house PZ1 [ ]), HRP-conjugated anti-rabbit and anti-mouse secondary antibodies (1:5000, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Western Blot, Expressing, MANN-WHITNEY, Staining

Journal: Scientific Reports

Article Title: Quantitative analysis of Gria1 , Gria2 , Dlg1 and Dlg4 expression levels in hippocampus following forced swim stress in mice

doi: 10.1038/s41598-019-50689-w

Figure Lengend Snippet: Exposure to the forced swim test for two consecutive days drastically reduces Dlg4 transcription. Hippocampal expression of Dlg1 , Dlg4 , Gria1 , and Gria2 mRNA (coding for SAP97, PSD-95, GluA1, and GluA2 respectively) relative to two reference genes ( Pgk1 and B2m ) after one (FST1; N = 8) or two (FST2; N = 7) swim test exposures relative to untreated wild type mice (CTRL; N = 9) (see Table and Fig. for details on primer pairs). Red circles indicate individual data points. Asterisks indicate statistical significance (Holm-Bonferroni corrected) compared to CTRL mice: ***P < 0.001 and n.s. indicates lack of significance compared to CTRL.

Article Snippet: For staining against SAP97: mouse anti-SAP97 1:500 (NeuroMab K64/15, Lot#75-030), mouse anti-GAPDH 1:10,000 (Sigma-Aldrich G8795, Lot#010M4814), mouse anti-β-actin 1:1,000 (Santa-Cruz sc-47778, Lot#C1510).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Quantitative analysis of Gria1 , Gria2 , Dlg1 and Dlg4 expression levels in hippocampus following forced swim stress in mice

doi: 10.1038/s41598-019-50689-w

Figure Lengend Snippet: Forced swim stress does not significantly alter protein levels in hippocampus. ( A ) Hippocampal SAP97, PSD-95, and GluA1 levels relative to β-actin and Gapdh after one (FST1; N = 3 for SAP97 homogenates, N = 10 for all others) or two (FST2; N = 7 for SAP97 synaptoneurosomes, N = 8 for SAP97 homogenates, N = 11 for all others) swim test exposures relative to untreated wild type mice (CTRL; N = 7 for SAP97 homogenates, N = 9 for all others). ( B ) Protein levels (relative to β-actin and Gapdh) in synaptoneurosomal fractions prepared from hippocampal homogenates (CTRL, N = 9; FST1, N = 10; FST2; N = 7–11). Note that differences in sample size are a result of samples being removed when the number of available replicates was below 3. Red circles indicate individual data points. Lack of significance compared to CTRL is indicated by n.s.

Article Snippet: For staining against SAP97: mouse anti-SAP97 1:500 (NeuroMab K64/15, Lot#75-030), mouse anti-GAPDH 1:10,000 (Sigma-Aldrich G8795, Lot#010M4814), mouse anti-β-actin 1:1,000 (Santa-Cruz sc-47778, Lot#C1510).

Techniques: