Anti-U1-C antibody [4H12]

• IP

Unknown

Immunoprecipitation - Anti-U1-C antibody [4H12] (AB122901)

Immunoprecipitation of U1-C with ab122901.
Lane 1 : no extract.
Lane 2 : HeLa total cell extract.
HL and LC : heavy and light chains.

All lanes:

Immunoprecipitation - Anti-U1-C antibody [4H12] (ab122901)

Predicted band size: 17 kDa

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• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-U1-C antibody [4H12] (AB122901)

ab122901 staining U1-C in Mouse L929 cells by Immunoflourescence (green). Right image shows the cells DNA stained with Hoechst.

• WB

CiteAb

Western blot - Anti-U1-C antibody [4H12] (AB122901)

Western Blotting using Anti-U1-C antibody [4H12], ab122901. Publication image from Pagani, F. et al., 2016, Nat Commun, 27041075. Legend direct from paper.

ExSpeU1s SM25 mutants with defective protein binding.(a) Schematic representation of RNA secondary structures and sequences of SM25 ExSpeU1 mutants. Highlighted modified nucleotides are shown in grey. (b) Expression levels, quality and nuclear distribution of the ExSpeU1 mutant RNAs. Representative northern blot analysis of ExSpeU1 mutants in total and nuclear RNA fractions. RNA was probed for ExSpeU1 SM25 and for U6 and the histograms below each northern blot show the ExSpeU1 SM25 variants' expression levels relative to U6. Values are the mean±s.e.m. of four independent experiments; *P≤0.05, **P≤0.01, ***P≤0.001. (c,d) Purity of total (T) and nuclear (N) fractions assessed by WB using Human Nuclear Matrix Protein p84, tubulin Ab and by EtBr staining of sRNAs loaded on 8% urea– polyacrylamide gel electrophoresis gel. Transfer RNA (tRNA) is missing in the nuclear fraction. (e) Affinity purification of ExSpeU1 SM25 mutants. Nuclear extracts (NE) from Hek293 cells transfected with the indicated constructs were incubated with the SM25 biotinylated oligonucleotide. Affinity-purified snRNPs were analysed by WB using antibodies against U1–70K, U1A and U1C. (f) RNA-IP analysis of SM25 ExSpeU1 variants. Hek293 NE from cells transfected with ExSpeU1 SM25 variants or not trasfected cells were incubated with antibodies against U1A, 70K, U1C and Sm proteins. RNAs purified from the RIP complexes were analysed by northern blotting. Pulled-down complexes were also analysed by northern blotting with U1 wild-type probe (Supplementary Fig. 9).

false

• WB

CiteAb

Western blot - Anti-U1-C antibody [4H12] (AB122901)

Western Blotting using Anti-U1-C antibody [4H12], ab122901. Publication image from Pagani, F. et al., 2016, Nat Commun, 27041075. Legend direct from paper.

Protein composition of ExSpeU1s and endogenous U1 snRNP.(a) Schematic representation of U1 snRNA secondary structure and associated proteins along with the RNA oligonucleotides used in affinity purification, RIP and EMSA. (b) Affinity purified CF11, SM25 and FIX9 ExSpeU1s contain 70K, U1A and U1C. Nuclear extracts (NE) from Hek293 cells, transfected with the indicated ExSpeU1s, or not transfected cells were incubated with the corresponding biotinylated 2′-O-methyl-RNA oligonucleotides. Affinity-purified snRNPs were analysed by western blotting using antibodies against U1–70K, U1A and U1C. (c) RNA-immunoprecipitation analysis of SM25 ExSpeU1. Hek293 NEs from cells transfected with ExSpeU1 SM25 or not transfected cells were incubated with antibodies against U1A, 70K and U1C. RNAs and proteins purified from the RIP complexes were analysed by northern and western blotting, respectively with the indicated probes/antibodies. Mock Ab corresponds to anti-tubulin. (d) CF11 ExSpeU1 snRNP has the same electrophoretic mobility as normal U1 in EMSA. Radiolabelled RNA oligonucleotides complementary to normal U1 (lanes 1–5) or ExSpeU1 CF11 (lanes 6–10) were incubated with nuclear extracts transfected with ExSpe CF11 or mock (not transfected cells). Addition of the indicated antibodies super-shifted the complexes in U1wt and ExSpe CF11. Control EMSA experiments are shown in Supplementary Fig. 5.

false

• WB

CiteAb

Western blot - Anti-U1-C antibody [4H12] (AB122901)

Western Blotting using Anti-U1-C antibody [4H12], ab122901. Publication image from Pagani, F. et al., 2016, Nat Commun, 27041075. Legend direct from paper.

Protein composition of ExSpeU1s and endogenous U1 snRNP.(a) Schematic representation of U1 snRNA secondary structure and associated proteins along with the RNA oligonucleotides used in affinity purification, RIP and EMSA. (b) Affinity purified CF11, SM25 and FIX9 ExSpeU1s contain 70K, U1A and U1C. Nuclear extracts (NE) from Hek293 cells, transfected with the indicated ExSpeU1s, or not transfected cells were incubated with the corresponding biotinylated 2′-O-methyl-RNA oligonucleotides. Affinity-purified snRNPs were analysed by western blotting using antibodies against U1–70K, U1A and U1C. (c) RNA-immunoprecipitation analysis of SM25 ExSpeU1. Hek293 NEs from cells transfected with ExSpeU1 SM25 or not transfected cells were incubated with antibodies against U1A, 70K and U1C. RNAs and proteins purified from the RIP complexes were analysed by northern and western blotting, respectively with the indicated probes/antibodies. Mock Ab corresponds to anti-tubulin. (d) CF11 ExSpeU1 snRNP has the same electrophoretic mobility as normal U1 in EMSA. Radiolabelled RNA oligonucleotides complementary to normal U1 (lanes 1–5) or ExSpeU1 CF11 (lanes 6–10) were incubated with nuclear extracts transfected with ExSpe CF11 or mock (not transfected cells). Addition of the indicated antibodies super-shifted the complexes in U1wt and ExSpe CF11. Control EMSA experiments are shown in Supplementary Fig. 5.

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