Antibody Generation and Characterization for Western Blot and Immunofluorescence

The anti-PARN antibody for western blot analysis was raised against purified, recombinant His-PARN and affinity purified with the antigen coupled to SulfoLink beads (Thermo Fisher Scientific). The anti-C21orf70 antibody was raised against a C-terminal peptide (C-GQTLARQMQLEDGGQL) and purified as described above. The antibody against RPL23a has been raised as previously described (40 (link)). The antibodies against DIM2, ENP1, NOB1, RPS3 (34 (link)), NOC4L, RRP12, RPS19 (29 (link)) and TSR1 (41 (link)) have been described previously. Anti-PARN for immunofluorescence was purchased from Abcam (ab89831), anti-β-actin and anti-α-tubulin from Sigma Aldrich (A1987 and T5168), anti-CK1δ and anti-HNRNPC from Abcam (AF12G4 and ab10294), anti-CK1ε (610445), anti-HA from Covance (MMS-101P) and anti-RPL10 from Santa Cruz Biotechnologies, Inc. (sc-798). Secondary antibodies for immunofluorescence were purchased from Invitrogen.

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Montellese C., Montel-Lehry N., Henras A.K., Kutay U., Gleizes P.E, & O’Donohue M.F. (2017). Poly(A)-specific ribonuclease is a nuclear ribosome biogenesis factor involved in human 18S rRNA maturation. Nucleic Acids Research, 45(11), 6822-6836.

Publication 2017

SulfoLink beads ab89831 anti-β-actin anti-α-tubulin

Actin Anti antibody Antibodies Antibody Antigen Immunofluorescence Peptide c Rpl10 Rps19 Rps3 Rrp12 Western blot analysis α tubulin

Corresponding Organization : Centre National de la Recherche Scientifique

Other organizations : ETH Zurich

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Variable analysis

independent variables

Purified, recombinant His-PARN protein used to raise anti-PARN antibody
C-terminal peptide (C-GQTLARQMQLEDGGQL) used to raise anti-C21orf70 antibody

dependent variables

Expression levels of PARN, C21orf70, RPL23a, DIM2, ENP1, NOB1, RPS3, NOC4L, RRP12, RPS19, and TSR1 proteins (as measured by western blot analysis)

control variables

β-actin
α-tubulin
CK1δ
HNRNPC
CK1ε
RPL10

positive controls

None specified

negative controls

None specified

Annotations

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