Anti-RALY antibody [EPR10121]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RALY antibody [EPR10121] (AB170105)

Immunohistochemical analysis of paraffin embedded Human heart tissue labeling RALY with ab170105 at a 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RALY antibody [EPR10121] (AB170105)

Immunohistochemical analysis of paraffin embedded Human brain tissue labeling RALY with ab170105 at a 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RALY antibody [EPR10121] (AB170105)

Immunofluorescence analysis of HepG2 cells labeling RALY with ab170105 at a 1/250 dilution.

• WB

Unknown

Western blot - Anti-RALY antibody [EPR10121] (AB170105)

All lanes:

Western blot - Anti-RALY antibody [EPR10121] (ab170105) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

A549 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 32 kDa

false

• WB

CiteAb

Western blot - Anti-RALY antibody [EPR10121] (AB170105)

Western Blotting using Anti-RALY antibody [EPR10121], ab170105. Publication image from Gorospe, M. et al., 2020, Nat Commun, 32958772. Legend direct from paper.

lincNORS regulates cholesterol synthesis via interaction with RALY.a The level of hnRNPC, RALY, TRIM28, SREBP2, and MS2-GST in the MS2 pulldown material and input lysate was detected by western blot. The data presented are a representative image from two to three independent experiments. b Subcellular location of RALY was detected by western blot. Tubulin, Lamin-B1, and acetyl-histone H3 were included as controls for cytoplasmic, nuclear, and chromatin-associated fractions. A representative image from three independent experiments is shown. c RNA immunoprecipitation (RIP) analysis of RALY. The presence of lincNORS and 18s rRNA in the precipitated complex was detected by qPCR. Data represent mean ± SD from three biological replicates (two-sided Student’s t-test). d qPCR analysis of NORS, MSMO1, and SQLE expression in MCF-7 cells transfected with control, lincNORS siRNA, and/or RALY siRNA. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). RALY knockdown was confirmed by western blot in each experiment and a representative image is shown on the right. e Enrichment of RNA of cholesterol synthesis genes in RALY-containing immunoprecipitated complex. The barplot displays the mean fold enrichment ± S.E.M. from three independent RALY RIP-Seq vs control experiments in MCF7 cells. The statistical significance of the enrichment was determined with CuffDiff v2.2.1 (*FDR < 0.05, **FDR < 0.01, ***FDR < 0.001). Source data are provided as a Source Data file. f Enrichment of MSMO1, SQLE, H1FX, and GAPDH in RALY-containing immunoprecipitated complex after lincNORS or hnRNPC knockdown was detected by qPCR. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test).

false

• WB

CiteAb

Western blot - Anti-RALY antibody [EPR10121] (AB170105)

Western Blotting using Anti-RALY antibody [EPR10121], ab170105. Publication image from Gorospe, M. et al., 2020, Nat Commun, 32958772. Legend direct from paper.

lincNORS regulates cholesterol synthesis via interaction with RALY.a The level of hnRNPC, RALY, TRIM28, SREBP2, and MS2-GST in the MS2 pulldown material and input lysate was detected by western blot. The data presented are a representative image from two to three independent experiments. b Subcellular location of RALY was detected by western blot. Tubulin, Lamin-B1, and acetyl-histone H3 were included as controls for cytoplasmic, nuclear, and chromatin-associated fractions. A representative image from three independent experiments is shown. c RNA immunoprecipitation (RIP) analysis of RALY. The presence of lincNORS and 18s rRNA in the precipitated complex was detected by qPCR. Data represent mean ± SD from three biological replicates (two-sided Student’s t-test). d qPCR analysis of NORS, MSMO1, and SQLE expression in MCF-7 cells transfected with control, lincNORS siRNA, and/or RALY siRNA. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). RALY knockdown was confirmed by western blot in each experiment and a representative image is shown on the right. e Enrichment of RNA of cholesterol synthesis genes in RALY-containing immunoprecipitated complex. The barplot displays the mean fold enrichment ± S.E.M. from three independent RALY RIP-Seq vs control experiments in MCF7 cells. The statistical significance of the enrichment was determined with CuffDiff v2.2.1 (*FDR < 0.05, **FDR < 0.01, ***FDR < 0.001). Source data are provided as a Source Data file. f Enrichment of MSMO1, SQLE, H1FX, and GAPDH in RALY-containing immunoprecipitated complex after lincNORS or hnRNPC knockdown was detected by qPCR. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test).

false

• WB

CiteAb

Western blot - Anti-RALY antibody [EPR10121] (AB170105)

Western Blotting using Anti-RALY antibody [EPR10121], ab170105. Publication image from Gorospe, M. et al., 2020, Nat Commun, 32958772. Legend direct from paper.

lincNORS regulates cholesterol synthesis via interaction with RALY.a The level of hnRNPC, RALY, TRIM28, SREBP2, and MS2-GST in the MS2 pulldown material and input lysate was detected by western blot. The data presented are a representative image from two to three independent experiments. b Subcellular location of RALY was detected by western blot. Tubulin, Lamin-B1, and acetyl-histone H3 were included as controls for cytoplasmic, nuclear, and chromatin-associated fractions. A representative image from three independent experiments is shown. c RNA immunoprecipitation (RIP) analysis of RALY. The presence of lincNORS and 18s rRNA in the precipitated complex was detected by qPCR. Data represent mean ± SD from three biological replicates (two-sided Student’s t-test). d qPCR analysis of NORS, MSMO1, and SQLE expression in MCF-7 cells transfected with control, lincNORS siRNA, and/or RALY siRNA. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). RALY knockdown was confirmed by western blot in each experiment and a representative image is shown on the right. e Enrichment of RNA of cholesterol synthesis genes in RALY-containing immunoprecipitated complex. The barplot displays the mean fold enrichment ± S.E.M. from three independent RALY RIP-Seq vs control experiments in MCF7 cells. The statistical significance of the enrichment was determined with CuffDiff v2.2.1 (*FDR < 0.05, **FDR < 0.01, ***FDR < 0.001). Source data are provided as a Source Data file. f Enrichment of MSMO1, SQLE, H1FX, and GAPDH in RALY-containing immunoprecipitated complex after lincNORS or hnRNPC knockdown was detected by qPCR. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test).

false

• WB

CiteAb

Western blot - Anti-RALY antibody [EPR10121] (AB170105)

Western Blotting using Anti-RALY antibody [EPR10121], ab170105. Publication image from Gorospe, M. et al., 2020, Nat Commun, 32958772. Legend direct from paper.

lincNORS regulates cholesterol synthesis via interaction with RALY.a The level of hnRNPC, RALY, TRIM28, SREBP2, and MS2-GST in the MS2 pulldown material and input lysate was detected by western blot. The data presented are a representative image from two to three independent experiments. b Subcellular location of RALY was detected by western blot. Tubulin, Lamin-B1, and acetyl-histone H3 were included as controls for cytoplasmic, nuclear, and chromatin-associated fractions. A representative image from three independent experiments is shown. c RNA immunoprecipitation (RIP) analysis of RALY. The presence of lincNORS and 18s rRNA in the precipitated complex was detected by qPCR. Data represent mean ± SD from three biological replicates (two-sided Student’s t-test). d qPCR analysis of NORS, MSMO1, and SQLE expression in MCF-7 cells transfected with control, lincNORS siRNA, and/or RALY siRNA. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). RALY knockdown was confirmed by western blot in each experiment and a representative image is shown on the right. e Enrichment of RNA of cholesterol synthesis genes in RALY-containing immunoprecipitated complex. The barplot displays the mean fold enrichment ± S.E.M. from three independent RALY RIP-Seq vs control experiments in MCF7 cells. The statistical significance of the enrichment was determined with CuffDiff v2.2.1 (*FDR < 0.05, **FDR < 0.01, ***FDR < 0.001). Source data are provided as a Source Data file. f Enrichment of MSMO1, SQLE, H1FX, and GAPDH in RALY-containing immunoprecipitated complex after lincNORS or hnRNPC knockdown was detected by qPCR. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test).

false