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Cfim59

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CFIm59 is a lab equipment product that plays a core role in the cellular process of pre-mRNA cleavage and polyadenylation. It functions as a component of the cleavage factor I (CFI) complex, which is involved in the regulation of gene expression by influencing the 3' end processing of mRNA transcripts.

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Lab products found in correlation

Cfim68, by Abcam (2 mentions) α tubulin, by Merck Group (1 mentions) U2af65, by Santa Cruz Biotechnology (1 mentions) β actin, by Merck Group (1 mentions) Flag m2, by Merck Group (1 mentions) Chemidoc mp system, by Bio-Rad (1 mentions)

2 protocols using cfim59

1

Protein Extraction and Immunoblotting Protocol

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Protein extracts were prepared by lysis in Laemmli buffer [62.5 mM Tris-HCl (pH 6.8), 5% 2-mercaptoethanol, 10% glycerol, 2% SDS, 0.002% bromophenol blue]. Blots were probed with the following antibodies: U2AF65 (Santa Cruz Biotechnology, MC3), MYC (Roche, 11-667-149-001), β-actin (Sigma, AC74), FLAG M2 (Sigma, F1804), ATG7 (ProSci, 3615), CFIm59 (Bethyl Laboratories, A301-359A), CFIm68 (Abcam, ab175237), LC3B (Cell Signaling Technology, #3868), p62 (Santa Cruz Biotechnology, sc-28359), U2AF35 (Proteintech, 10334-1-AP) and α-tubulin (Sigma, B5-1-2). Immunoblots were visualized by a ChemiDoc™ MP System (Bio-Rad).
Park S.M., Ou J., Chamberlain L., Simone T.M., Yang H., Virbasius C.M., Ali A.M., Zhu L.J., Mukherjee S., Raza A, & Green M.R. (2016). U2AF35(S34F) Promotes Transformation by Directing Aberrant ATG7 Pre-mRNA 3’ End Formation. Molecular cell, 62(4), 479-490.
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2

RNA Immunoprecipitation Protocol

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RNA immunoprecipitation was performed as described in the Supplemental Experimental Procedures, using the following antibodies: CFIm25 (Santa Cruz, sc-81109), CFIm59 (Bethyl Laboratories, A301-360A) and CFIm68 (Abcam, ab175237) were used for RNA immunoprecipitation and mouse IgG (Abcam, ab18413) or rabbit IgG (Abcam, ab46540) as a negative control. 1% of the DNase-treated extract was taken for an input fraction. The qRT-PCR levels of the IP samples were normalized against the input of the same samples and calibrated to an IgG control followed by normalization to an irrelevant region in the 18S rRNA. Fold enrichment was calculated by setting the IgG control IP sample to a value of 1.
Park S.M., Ou J., Chamberlain L., Simone T.M., Yang H., Virbasius C.M., Ali A.M., Zhu L.J., Mukherjee S., Raza A, & Green M.R. (2016). U2AF35(S34F) Promotes Transformation by Directing Aberrant ATG7 Pre-mRNA 3’ End Formation. Molecular cell, 62(4), 479-490.
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