The following antibodies were used in this study: hnRNPF/H (sc-32310, RRID:AB_2248257), Splicing factor, proline- and glutamine-rich (SFPQ) (sc-374502, RRID:AB_10989589), non-POU domain-containing octamer-binding protein (NONO) (sc-166702, RRID:AB_2152178), hnRNPQ (sc-56703, RRID:AB_2200715), hnRNPA2/B1 (sc-374053, RRID:AB_10947257), and glutathione S-transferase (GST) (sc-138, RRID:AB_627677) from Santa Cruz Biotechnology (Dallas, TX); hnRNPM (A500–011A, RRID:AB_11125542) from Bethyl Laboratories (Montgomery, TX); horseradish peroxidase (HRP)-conjugated FLAG (A8592, RRID:AB_439702) from Sigma-Aldrich (St. Louis, MO); HA (11867423001, RRID:AB_390918) and HRP-conjugated HA (12013819001, RRID:AB_390917) from Roche Diagnostics (Basel, Swiss); monoclonal TDP-43 (89789, RRID:AB_2800143) and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (2118, RRID:AB_561053) from Cell Signaling Technology (Danvers, MA); β-Tubulin (014–25041, RRID:AB_2650453) from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan); polyclonal TDP-43 (12892–1-AP, RRID:AB_2200505) and polyclonal Matrin3 (MATR3) (12202–2-AP, RRID:AB_2281752) from Proteintech Group (Rosemont, IL); HRP-conjugated goat anti-rabbit IgG (H + L) secondary antibody (170–6515, RRID:AB_11125142) and HRP-conjugated goat anti-mouse IgG (H + L) secondary antibody (170–6516, RRID:AB_11125547) from Bio-Rad Laboratories (Hercules, CA).
Hnrnpf h
Manufactured by Santa Cruz Biotechnology
The HnRNPF/H product is a recombinant protein that represents the heterogeneous nuclear ribonucleoproteins F and H. These proteins play a role in pre-mRNA processing and other cellular processes. This product is intended for research use only.
Automatically generated - may contain errors
Lab products found in correlation
Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti rabbit igg h l secondary antibody, by Proteintech (1 mentions)
Hnrnpa2 b1, by Santa Cruz Biotechnology (1 mentions)
β tubulin, by Fujifilm (1 mentions)
Tdp 43, by Cell Signaling Technology (1 mentions)
Taqman microrna assay, by Thermo Fisher Scientific (1 mentions)
2 protocols using hnrnpf h
1
Antibody panel for RNA-binding proteins
2
Quantification of tRF and U7 snRNA in ES Cells
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RNA immunoprecipitations were done using a 10-cm dish of mouse E14 ES cells. Five-hundred micrograms of lysate was incubated with either 10 μg of hnRNP F/H (Santa Cruz Biotechnology) antibody or 10 μg of mouse IgG-conjugated Protein G Dynabeads overnight in the cold room. The beads were then washed sequentially for five washes with Millipore EZ-Magna-RIP wash buffer (#17-701). RNA was then isolated from the beads using Trizol and precipitated using isopropanol following standard procedures. Purified RNA quality was checked using bioanalyzer. tRF and U7 snRNA quantification was performed using custom designed TaqMan microRNA assays according to manufacturer's recommended protocols (Applied Biosystems). Ten nanograms of input RNA, RNA-IP, and control IP RNA was reverse transcribed using the TaqMan microRNA reverse transcription kit. qRT-PCR was performed in 15-µL reactions using TaqMan Universal PCR Master Mix, following standard program (10 min at 95°C, then 15 sec at 95°C, and 1 min at 60°C for 40 cycles).
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