Staufen Antibody

Western Blot: Staufen Antibody [NBP1-33202]

Western Blot: Staufen Antibody [NBP1-33202] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with Staufen antibody diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

Immunocytochemistry/ Immunofluorescence: Staufen Antibody [NBP1-33202]

Immunocytochemistry/Immunofluorescence: Staufen Antibody [NBP1-33202] - DIV9 rat E18 primary hippocampal neuron cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Staufen stained by Staufen antibody diluted at 1:500.Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody [11710] diluted at 1:500. Blue: Fluoroshield with DAPI.

Immunoprecipitation: Staufen Antibody [NBP1-33202]

Immunoprecipitation: Staufen Antibody [NBP1-33202] - Staufen antibody staining 30 ug whole cell lysate of Lane A: 293 (input), Lane B: control rabbit IgG-IP, Lane C: Stau1-IP, Lane D: Post-IP lysate from control rabbit IgG-IP, Lane E:Post-IP lysate from Stau1-IP; diluted at 1:3000.

Western Blot: Staufen Antibody [NBP1-33202] -

Staufen1 protein but not mRNA steady-state levels are increased in neurodegenerative disease cells & tissues. Western blot analysis of SCA2- FBs (a) & LBCs (b) show increased STAU1 levels compared with normal controls. DDX6 levels are unchanged. HD & SCA3 patient (polyQ expanded) FBs were used as additional controls. Four normal & five SCA2 FBs, & two normal & three SCA2 LBCs were used. c, d Western blot analyses of ATXN2Q127 (c) & BAC-Q72 (d) mouse cerebellar extracts (24 weeks of age) showing increased Stau1 levels compared with wild-type or BAC-Q22 controls (n = 2–3 animals per group). e Western blot of FB extracts from an ALS patient with the TDP-43G298S mutation show increased STAU1 levels. beta-Actin was used as loading control & representative blots of three independent experiments are shown. f–hSTAU1 RNA levels are unaltered in SCA2 & ALS cells & SCA2 mice. qRT-PCR analyses of STAU1 mRNA in SCA2 FBs & ALS FB with TDP-43G298S mutation (f) or SCA2 LBCs (g). h qRT-PCR analyses of cerebellar RNAs from ATXN2Q127 & BAC-Q72 mice compared to wild-type littermates (24 weeks of age; n = animals per group). Gene expression levels were normalized to Actb Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Silencing of STAU1 mitigates SCA2 phenotypes. aStau1 haploinsufficiency improves abnormal motor behavior of ATXN2Q127 mice as determined by rotarod behavior at 8, 12, 16, & 20 weeks of age. ATXN2Q127;Stau1+/- mice (green) have improved rotarod performance compared with ATXN2Q127 littermates (red) starting at 12 weeks of age. Note that Stau1 haploinsufficiency (orange) by itself does not alter motor function; n = 9–15 mice per group. Values shown are mean ± SE. Significance was determined using generalized estimating equations (GEE). NS, nonsignificant, *P < 0.05, **P < 0.01. b, c Reduction of Stau1 in vivo improves levels of key cerebellar proteins towards normalization. b Western blotting of cerebellar extracts from ATXN2Q127;Stau1+/- mice showing improvement of protein levels for Calb1, Pcp2, Rgs8, Pcp4, Homer3, & Fam107b towards normalization. Each lane represents cerebellar extract from an individual mouse. beta-Actin is used as a loading control & the blots are from three replicate experiments. c Quantitative analysis of western blots shown in b. Data are mean ± SD, **P < 0.01, ***P < 0.001, Student t-test. d Combined immunostaining of ATXN2 (red) & Stau1 (green) of cerebellar sections from ATXN2Q127 & crossed ATXN2Q127;Stau1+/- mice (34 weeks of age) demonstrating reduced ATXN2-Stau1 aggregates in crossed ATXN2Q127;Stau1+/- mice. Scale bar, 30 µM. e Model for STAU1 in the pathology of SCA2 & other neurodegenerative diseases Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Staufen1 protein but not mRNA steady-state levels are increased in neurodegenerative disease cells & tissues. Western blot analysis of SCA2- FBs (a) & LBCs (b) show increased STAU1 levels compared with normal controls. DDX6 levels are unchanged. HD & SCA3 patient (polyQ expanded) FBs were used as additional controls. Four normal & five SCA2 FBs, & two normal & three SCA2 LBCs were used. c, d Western blot analyses of ATXN2Q127 (c) & BAC-Q72 (d) mouse cerebellar extracts (24 weeks of age) showing increased Stau1 levels compared with wild-type or BAC-Q22 controls (n = 2–3 animals per group). e Western blot of FB extracts from an ALS patient with the TDP-43G298S mutation show increased STAU1 levels. beta-Actin was used as loading control & representative blots of three independent experiments are shown. f–hSTAU1 RNA levels are unaltered in SCA2 & ALS cells & SCA2 mice. qRT-PCR analyses of STAU1 mRNA in SCA2 FBs & ALS FB with TDP-43G298S mutation (f) or SCA2 LBCs (g). h qRT-PCR analyses of cerebellar RNAs from ATXN2Q127 & BAC-Q72 mice compared to wild-type littermates (24 weeks of age; n = animals per group). Gene expression levels were normalized to Actb Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Staufen1 protein but not mRNA steady-state levels are increased in neurodegenerative disease cells & tissues. Western blot analysis of SCA2- FBs (a) & LBCs (b) show increased STAU1 levels compared with normal controls. DDX6 levels are unchanged. HD & SCA3 patient (polyQ expanded) FBs were used as additional controls. Four normal & five SCA2 FBs, & two normal & three SCA2 LBCs were used. c, d Western blot analyses of ATXN2Q127 (c) & BAC-Q72 (d) mouse cerebellar extracts (24 weeks of age) showing increased Stau1 levels compared with wild-type or BAC-Q22 controls (n = 2–3 animals per group). e Western blot of FB extracts from an ALS patient with the TDP-43G298S mutation show increased STAU1 levels. beta-Actin was used as loading control & representative blots of three independent experiments are shown. f–hSTAU1 RNA levels are unaltered in SCA2 & ALS cells & SCA2 mice. qRT-PCR analyses of STAU1 mRNA in SCA2 FBs & ALS FB with TDP-43G298S mutation (f) or SCA2 LBCs (g). h qRT-PCR analyses of cerebellar RNAs from ATXN2Q127 & BAC-Q72 mice compared to wild-type littermates (24 weeks of age; n = animals per group). Gene expression levels were normalized to Actb Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Co-localization of Staufen1 with ATXN2 in stress-granule-like structures. a ATXN2 & STAU1 co-localize in SGs. Immunostaining of HEK-293 cells with antibodies against ATXN2, STAU1, & TIA-1 show co-localization of STAU1 with ATXN2 & TIA-1 in SGs during stress (0.5 mM sodium arsenite for 15 & 30 min) that are not present in the absence of stress. Scale bar, 10 µM. b Constitutively present SG-like structures positive for both ATXN2 & STAU1 in SCA2 FBs, but not in normal FBs (white arrows). Cells were stained with antibodies to ATXN2 & STAU1. Scale bar, 100 µM. c Increased numbers of aggregates in SCA2 FBs at 37 °C. Aggregates > 4 pixels per cell positive for both ATXN2 & STAU1 are shown. One-hundred normal & 96 SCA2-FBs were used for analyses. Data are mean ± SD, **P < 0.01, Student t-test. d Histograms representing the quantities & sizes of ATXN2-STAU1 co-localized granules in normal vs. SCA2 FBs. e In vivo co-localization of ATXN2 (green) with Stau1 (red) to aggregates in cerebellar PCs of 24-weeks-old ATXN2Q127 mice (white arrows). Scale bar, 30 µM. f Co-localized ATXN2 (red) & STAU1 (green) to aggregates (white arrows) in cerebellar PCs from a human SCA2 brain, that are absent in an unaffected control. Scale bar, 30 µM. g Immunoprecipitation of ATXN2 with STAU1. Non-RNase A or RNase A treated HEK-293 cell extracts expressing Flag-tagged ATXN2-(Q22 or Q108) were subjected to immunoprecipitation with Flag mAb beads & analyzed by western blotting. STAU1 shows RNA-dependent interactions with wild-type & mutant ATXN2. ATXN2 also co-immunoprecipitates with DDX6 & PABPC1, known ATXN2 interactors. Representative blots of three independent experiments are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Co-localization of Staufen1 with ATXN2 in stress-granule-like structures. a ATXN2 & STAU1 co-localize in SGs. Immunostaining of HEK-293 cells with antibodies against ATXN2, STAU1, & TIA-1 show co-localization of STAU1 with ATXN2 & TIA-1 in SGs during stress (0.5 mM sodium arsenite for 15 & 30 min) that are not present in the absence of stress. Scale bar, 10 µM. b Constitutively present SG-like structures positive for both ATXN2 & STAU1 in SCA2 FBs, but not in normal FBs (white arrows). Cells were stained with antibodies to ATXN2 & STAU1. Scale bar, 100 µM. c Increased numbers of aggregates in SCA2 FBs at 37 °C. Aggregates > 4 pixels per cell positive for both ATXN2 & STAU1 are shown. One-hundred normal & 96 SCA2-FBs were used for analyses. Data are mean ± SD, **P < 0.01, Student t-test. d Histograms representing the quantities & sizes of ATXN2-STAU1 co-localized granules in normal vs. SCA2 FBs. e In vivo co-localization of ATXN2 (green) with Stau1 (red) to aggregates in cerebellar PCs of 24-weeks-old ATXN2Q127 mice (white arrows). Scale bar, 30 µM. f Co-localized ATXN2 (red) & STAU1 (green) to aggregates (white arrows) in cerebellar PCs from a human SCA2 brain, that are absent in an unaffected control. Scale bar, 30 µM. g Immunoprecipitation of ATXN2 with STAU1. Non-RNase A or RNase A treated HEK-293 cell extracts expressing Flag-tagged ATXN2-(Q22 or Q108) were subjected to immunoprecipitation with Flag mAb beads & analyzed by western blotting. STAU1 shows RNA-dependent interactions with wild-type & mutant ATXN2. ATXN2 also co-immunoprecipitates with DDX6 & PABPC1, known ATXN2 interactors. Representative blots of three independent experiments are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Silencing of STAU1 mitigates SCA2 phenotypes. aStau1 haploinsufficiency improves abnormal motor behavior of ATXN2Q127 mice as determined by rotarod behavior at 8, 12, 16, & 20 weeks of age. ATXN2Q127;Stau1+/- mice (green) have improved rotarod performance compared with ATXN2Q127 littermates (red) starting at 12 weeks of age. Note that Stau1 haploinsufficiency (orange) by itself does not alter motor function; n = 9–15 mice per group. Values shown are mean ± SE. Significance was determined using generalized estimating equations (GEE). NS, nonsignificant, *P < 0.05, **P < 0.01. b, c Reduction of Stau1 in vivo improves levels of key cerebellar proteins towards normalization. b Western blotting of cerebellar extracts from ATXN2Q127;Stau1+/- mice showing improvement of protein levels for Calb1, Pcp2, Rgs8, Pcp4, Homer3, & Fam107b towards normalization. Each lane represents cerebellar extract from an individual mouse. beta-Actin is used as a loading control & the blots are from three replicate experiments. c Quantitative analysis of western blots shown in b. Data are mean ± SD, **P < 0.01, ***P < 0.001, Student t-test. d Combined immunostaining of ATXN2 (red) & Stau1 (green) of cerebellar sections from ATXN2Q127 & crossed ATXN2Q127;Stau1+/- mice (34 weeks of age) demonstrating reduced ATXN2-Stau1 aggregates in crossed ATXN2Q127;Stau1+/- mice. Scale bar, 30 µM. e Model for STAU1 in the pathology of SCA2 & other neurodegenerative diseases Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Staufen1 protein but not mRNA steady-state levels are increased in neurodegenerative disease cells & tissues. Western blot analysis of SCA2- FBs (a) & LBCs (b) show increased STAU1 levels compared with normal controls. DDX6 levels are unchanged. HD & SCA3 patient (polyQ expanded) FBs were used as additional controls. Four normal & five SCA2 FBs, & two normal & three SCA2 LBCs were used. c, d Western blot analyses of ATXN2Q127 (c) & BAC-Q72 (d) mouse cerebellar extracts (24 weeks of age) showing increased Stau1 levels compared with wild-type or BAC-Q22 controls (n = 2–3 animals per group). e Western blot of FB extracts from an ALS patient with the TDP-43G298S mutation show increased STAU1 levels. beta-Actin was used as loading control & representative blots of three independent experiments are shown. f–hSTAU1 RNA levels are unaltered in SCA2 & ALS cells & SCA2 mice. qRT-PCR analyses of STAU1 mRNA in SCA2 FBs & ALS FB with TDP-43G298S mutation (f) or SCA2 LBCs (g). h qRT-PCR analyses of cerebellar RNAs from ATXN2Q127 & BAC-Q72 mice compared to wild-type littermates (24 weeks of age; n = animals per group). Gene expression levels were normalized to Actb Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Staufen Antibody [NBP1-33202] -

Western Blot: Staufen Antibody [NBP1-33202] - Co-localization of Staufen1 with ATXN2 in stress-granule-like structures. a ATXN2 & STAU1 co-localize in SGs. Immunostaining of HEK-293 cells with antibodies against ATXN2, STAU1, & TIA-1 show co-localization of STAU1 with ATXN2 & TIA-1 in SGs during stress (0.5 mM sodium arsenite for 15 & 30 min) that are not present in the absence of stress. Scale bar, 10 µM. b Constitutively present SG-like structures positive for both ATXN2 & STAU1 in SCA2 FBs, but not in normal FBs (white arrows). Cells were stained with antibodies to ATXN2 & STAU1. Scale bar, 100 µM. c Increased numbers of aggregates in SCA2 FBs at 37 °C. Aggregates > 4 pixels per cell positive for both ATXN2 & STAU1 are shown. One-hundred normal & 96 SCA2-FBs were used for analyses. Data are mean ± SD, **P < 0.01, Student t-test. d Histograms representing the quantities & sizes of ATXN2-STAU1 co-localized granules in normal vs. SCA2 FBs. e In vivo co-localization of ATXN2 (green) with Stau1 (red) to aggregates in cerebellar PCs of 24-weeks-old ATXN2Q127 mice (white arrows). Scale bar, 30 µM. f Co-localized ATXN2 (red) & STAU1 (green) to aggregates (white arrows) in cerebellar PCs from a human SCA2 brain, that are absent in an unaffected control. Scale bar, 30 µM. g Immunoprecipitation of ATXN2 with STAU1. Non-RNase A or RNase A treated HEK-293 cell extracts expressing Flag-tagged ATXN2-(Q22 or Q108) were subjected to immunoprecipitation with Flag mAb beads & analyzed by western blotting. STAU1 shows RNA-dependent interactions with wild-type & mutant ATXN2. ATXN2 also co-immunoprecipitates with DDX6 & PABPC1, known ATXN2 interactors. Representative blots of three independent experiments are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30194296), licensed under a CC-BY license. Not internally tested by Novus Biologicals.