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rabbit polyclonal antibodies against cpsf5 (ABclonal Biotechnology)

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ABclonal Biotechnology rabbit polyclonal antibodies against cpsf5

<t>CPSF5</t> recruited by NP1 targets viral RNAs to further regulate the alternative RNA processing. ( A ) NP1 interacts with CPSF5, which was verified by performing an IP assay. Flag-tagged NP1 was transiently expressed in WRD cells and then purified by IP using anti-Flag antibodies and detected via MS (Table ) or western blotting. IgG was used as a negative control. ( B ) Southern blot assay showing MVC DNA replication in CPSF5 knockdown cells. Hirt DNA was extracted from MVC-infected cells with CPSF5 knockdown and subjected to Southern blot analysis at 48 h post-infection. ( C ) The expression of MVC protein was identified with CPSF5 knockdown. Western blot analysis of MVC protein expression by knocking down CPSF5 in WRD cells transfected with WT or mutant 3311m MVC infectious clone at 48 h; actin was used as a control. Relative intensity of VP2 versus actin was quantified using the ImageJ program. Data are means ± SDs ( n = 3). ***P ≤ 0.001, **P ≤ 0.01, unpaired Student’s t -test. ( D , E ) MVC RNA levels detected in CPSF5 knockdown cells. qRT-PCR was performed to determine the RNA levels from CPSF5 knockdown WRD cells transfected with infectious clone WT ( D ) or mutant 3311m ( E ) at 48 h by CPSF5, NP1, VP2 ORF primers, and GAPDH was used as a control. Data are means ± SEMs ( n = 3). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ns: not significant, unpaired Student’s t -tests. ( F , G ) The relationship between CPSF5 and the ac4C-modified residue in regulating RNA processing was detected by RPA. RPA of total RNA extracted from CPSF5-knockdown WRD cells transfected with WT or 3311m mutant MVC infectious clone at 48 h was performed using a 3D-probe ( F ) and (pA)p-probe ( G ). ( H , I ) Ability of CPSF5 binding to the targeted MVC RNA based on formaldehyde-RIP-qRT-PCR. Flag-tagged CPSF5 was expressed in WRD cells, and formaldehyde-cross-linking cell lysates were subjected to IP with IgG or anti-Flag antibodies ( H ). WT and 3311m mutant infectious clones were used to transfect WRD cells in which CPSF5 was knocked down, then the cells were subjected to IP with anti-CPSF5 antibodies. qRT-PCR was performed to quantify MVC RNA ( I ). IgG was used as a negative control. Unpaired student’s t -tests were performed, and the data are presented as the means ± SEMs ( n = 3). ***P ≤ 0.001.

Rabbit Polyclonal Antibodies Against Cpsf5, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+antibodies+against+cpsf5/pmc11959542-90-24-30?v=ABclonal+Biotechnology
Average 90 stars, based on 1 article reviews

rabbit polyclonal antibodies against cpsf5 - by Bioz Stars, 2026-07

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1) Product Images from "N4-acetylcytidine coordinates with NP1 and CPSF5 to facilitate alternative RNA processing during the replication of minute virus of canines"

Article Title: N4-acetylcytidine coordinates with NP1 and CPSF5 to facilitate alternative RNA processing during the replication of minute virus of canines

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkaf229

CPSF5 recruited by NP1 targets viral RNAs to further regulate the alternative RNA processing. ( A ) NP1 interacts with CPSF5, which was verified by performing an IP assay. Flag-tagged NP1 was transiently expressed in WRD cells and then purified by IP using anti-Flag antibodies and detected via MS (Table ) or western blotting. IgG was used as a negative control. ( B ) Southern blot assay showing MVC DNA replication in CPSF5 knockdown cells. Hirt DNA was extracted from MVC-infected cells with CPSF5 knockdown and subjected to Southern blot analysis at 48 h post-infection. ( C ) The expression of MVC protein was identified with CPSF5 knockdown. Western blot analysis of MVC protein expression by knocking down CPSF5 in WRD cells transfected with WT or mutant 3311m MVC infectious clone at 48 h; actin was used as a control. Relative intensity of VP2 versus actin was quantified using the ImageJ program. Data are means ± SDs ( n = 3). ***P ≤ 0.001, **P ≤ 0.01, unpaired Student’s t -test. ( D , E ) MVC RNA levels detected in CPSF5 knockdown cells. qRT-PCR was performed to determine the RNA levels from CPSF5 knockdown WRD cells transfected with infectious clone WT ( D ) or mutant 3311m ( E ) at 48 h by CPSF5, NP1, VP2 ORF primers, and GAPDH was used as a control. Data are means ± SEMs ( n = 3). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ns: not significant, unpaired Student’s t -tests. ( F , G ) The relationship between CPSF5 and the ac4C-modified residue in regulating RNA processing was detected by RPA. RPA of total RNA extracted from CPSF5-knockdown WRD cells transfected with WT or 3311m mutant MVC infectious clone at 48 h was performed using a 3D-probe ( F ) and (pA)p-probe ( G ). ( H , I ) Ability of CPSF5 binding to the targeted MVC RNA based on formaldehyde-RIP-qRT-PCR. Flag-tagged CPSF5 was expressed in WRD cells, and formaldehyde-cross-linking cell lysates were subjected to IP with IgG or anti-Flag antibodies ( H ). WT and 3311m mutant infectious clones were used to transfect WRD cells in which CPSF5 was knocked down, then the cells were subjected to IP with anti-CPSF5 antibodies. qRT-PCR was performed to quantify MVC RNA ( I ). IgG was used as a negative control. Unpaired student’s t -tests were performed, and the data are presented as the means ± SEMs ( n = 3). ***P ≤ 0.001.

Figure Legend Snippet: CPSF5 recruited by NP1 targets viral RNAs to further regulate the alternative RNA processing. ( A ) NP1 interacts with CPSF5, which was verified by performing an IP assay. Flag-tagged NP1 was transiently expressed in WRD cells and then purified by IP using anti-Flag antibodies and detected via MS (Table ) or western blotting. IgG was used as a negative control. ( B ) Southern blot assay showing MVC DNA replication in CPSF5 knockdown cells. Hirt DNA was extracted from MVC-infected cells with CPSF5 knockdown and subjected to Southern blot analysis at 48 h post-infection. ( C ) The expression of MVC protein was identified with CPSF5 knockdown. Western blot analysis of MVC protein expression by knocking down CPSF5 in WRD cells transfected with WT or mutant 3311m MVC infectious clone at 48 h; actin was used as a control. Relative intensity of VP2 versus actin was quantified using the ImageJ program. Data are means ± SDs ( n = 3). ***P ≤ 0.001, **P ≤ 0.01, unpaired Student’s t -test. ( D , E ) MVC RNA levels detected in CPSF5 knockdown cells. qRT-PCR was performed to determine the RNA levels from CPSF5 knockdown WRD cells transfected with infectious clone WT ( D ) or mutant 3311m ( E ) at 48 h by CPSF5, NP1, VP2 ORF primers, and GAPDH was used as a control. Data are means ± SEMs ( n = 3). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ns: not significant, unpaired Student’s t -tests. ( F , G ) The relationship between CPSF5 and the ac4C-modified residue in regulating RNA processing was detected by RPA. RPA of total RNA extracted from CPSF5-knockdown WRD cells transfected with WT or 3311m mutant MVC infectious clone at 48 h was performed using a 3D-probe ( F ) and (pA)p-probe ( G ). ( H , I ) Ability of CPSF5 binding to the targeted MVC RNA based on formaldehyde-RIP-qRT-PCR. Flag-tagged CPSF5 was expressed in WRD cells, and formaldehyde-cross-linking cell lysates were subjected to IP with IgG or anti-Flag antibodies ( H ). WT and 3311m mutant infectious clones were used to transfect WRD cells in which CPSF5 was knocked down, then the cells were subjected to IP with anti-CPSF5 antibodies. qRT-PCR was performed to quantify MVC RNA ( I ). IgG was used as a negative control. Unpaired student’s t -tests were performed, and the data are presented as the means ± SEMs ( n = 3). ***P ≤ 0.001.

Techniques Used: Purification, Western Blot, Negative Control, Southern Blot, Knockdown, Infection, Expressing, Transfection, Mutagenesis, Control, Quantitative RT-PCR, Modification, Residue, Binding Assay, Clone Assay

Figure Legend Snippet: MS data of NP1

Techniques Used:

CPSF5 mediated MVC RNA splicing is dependent on NP1 and ac4C modification at residue 3311. ( A , B ) The effect on CPSF5 binding to MVC RNA by increasing NP1 expression using formaldehyde-RIP-qRT-PCR. Vector or HA-NP1 and infectious clone WT or 3311m were co-transfected in WRD cells. Cell lysates cross-linked by formaldehyde were subjected to IP with IgG, anti-CPSF5 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). *** P ≤ 0.001, ns: not significant. ( C , D ) The effect on NP1 binding to MVC RNA by increasing CPSF5 expression using formaldehyde-RIP-qRT-PCR. Vector or Flag-CPSF5 and infectious clone WT or mutant 3311m were co-transfected in WRD cells. Cell lysates were subjected to IP with IgG, anti-NP1 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). ns: not significant. ( E ) Characterization of the cleavage and polyadenylation sites and downstream elements (DSEs) in (pA)p is shown at the top. The bottom panel shows a schematic representation of the RNA pulldown assay. The DNA template for T7 transcription consisted of three repeats of modified DSE. ( F , G ) Biotinylated RNA affinities of NP1 and CPSF5. HA-tagged NP1 or the vector was transiently expressed in WRD cells, and cell lysates were used to bind biotinylated RNA with or without the ac4C modification. In addition, purified NP1 or CPSF5 protein was mixed with RNA oligonucleotides with or without ac4C-modified RNA and analyzed via western blotting. ( H , I ) The binding analysis of CPSF5 or NP1 to oligo-RNA with or without the ac4C modification in the presence or absence of the NP1 ( H ) or CPSF5 ( I ) protein. ( J , K ) The binding analysis of NP1 and CPSF5 to ac4C-modified RNA. Equal amounts of purified CPSF5 ( J ) or NP1 ( K ) protein and increasing amounts of NP1 ( J ) or CPSF5 ( K ) protein were incubated with ac4C-modified RNA and analyzed via western blotting. Blots were detected using anti-CPSF5 or anti-NP1 antibodies.

Figure Legend Snippet: CPSF5 mediated MVC RNA splicing is dependent on NP1 and ac4C modification at residue 3311. ( A , B ) The effect on CPSF5 binding to MVC RNA by increasing NP1 expression using formaldehyde-RIP-qRT-PCR. Vector or HA-NP1 and infectious clone WT or 3311m were co-transfected in WRD cells. Cell lysates cross-linked by formaldehyde were subjected to IP with IgG, anti-CPSF5 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). *** P ≤ 0.001, ns: not significant. ( C , D ) The effect on NP1 binding to MVC RNA by increasing CPSF5 expression using formaldehyde-RIP-qRT-PCR. Vector or Flag-CPSF5 and infectious clone WT or mutant 3311m were co-transfected in WRD cells. Cell lysates were subjected to IP with IgG, anti-NP1 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). ns: not significant. ( E ) Characterization of the cleavage and polyadenylation sites and downstream elements (DSEs) in (pA)p is shown at the top. The bottom panel shows a schematic representation of the RNA pulldown assay. The DNA template for T7 transcription consisted of three repeats of modified DSE. ( F , G ) Biotinylated RNA affinities of NP1 and CPSF5. HA-tagged NP1 or the vector was transiently expressed in WRD cells, and cell lysates were used to bind biotinylated RNA with or without the ac4C modification. In addition, purified NP1 or CPSF5 protein was mixed with RNA oligonucleotides with or without ac4C-modified RNA and analyzed via western blotting. ( H , I ) The binding analysis of CPSF5 or NP1 to oligo-RNA with or without the ac4C modification in the presence or absence of the NP1 ( H ) or CPSF5 ( I ) protein. ( J , K ) The binding analysis of NP1 and CPSF5 to ac4C-modified RNA. Equal amounts of purified CPSF5 ( J ) or NP1 ( K ) protein and increasing amounts of NP1 ( J ) or CPSF5 ( K ) protein were incubated with ac4C-modified RNA and analyzed via western blotting. Blots were detected using anti-CPSF5 or anti-NP1 antibodies.

Techniques Used: Modification, Residue, Binding Assay, Expressing, Quantitative RT-PCR, Plasmid Preparation, Transfection, Negative Control, Mutagenesis, Purification, Western Blot, Incubation

Image Search Results

Journal: Nucleic Acids Research

Article Title: N4-acetylcytidine coordinates with NP1 and CPSF5 to facilitate alternative RNA processing during the replication of minute virus of canines

doi: 10.1093/nar/gkaf229

Figure Lengend Snippet: CPSF5 recruited by NP1 targets viral RNAs to further regulate the alternative RNA processing. ( A ) NP1 interacts with CPSF5, which was verified by performing an IP assay. Flag-tagged NP1 was transiently expressed in WRD cells and then purified by IP using anti-Flag antibodies and detected via MS (Table ) or western blotting. IgG was used as a negative control. ( B ) Southern blot assay showing MVC DNA replication in CPSF5 knockdown cells. Hirt DNA was extracted from MVC-infected cells with CPSF5 knockdown and subjected to Southern blot analysis at 48 h post-infection. ( C ) The expression of MVC protein was identified with CPSF5 knockdown. Western blot analysis of MVC protein expression by knocking down CPSF5 in WRD cells transfected with WT or mutant 3311m MVC infectious clone at 48 h; actin was used as a control. Relative intensity of VP2 versus actin was quantified using the ImageJ program. Data are means ± SDs ( n = 3). ***P ≤ 0.001, **P ≤ 0.01, unpaired Student’s t -test. ( D , E ) MVC RNA levels detected in CPSF5 knockdown cells. qRT-PCR was performed to determine the RNA levels from CPSF5 knockdown WRD cells transfected with infectious clone WT ( D ) or mutant 3311m ( E ) at 48 h by CPSF5, NP1, VP2 ORF primers, and GAPDH was used as a control. Data are means ± SEMs ( n = 3). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ns: not significant, unpaired Student’s t -tests. ( F , G ) The relationship between CPSF5 and the ac4C-modified residue in regulating RNA processing was detected by RPA. RPA of total RNA extracted from CPSF5-knockdown WRD cells transfected with WT or 3311m mutant MVC infectious clone at 48 h was performed using a 3D-probe ( F ) and (pA)p-probe ( G ). ( H , I ) Ability of CPSF5 binding to the targeted MVC RNA based on formaldehyde-RIP-qRT-PCR. Flag-tagged CPSF5 was expressed in WRD cells, and formaldehyde-cross-linking cell lysates were subjected to IP with IgG or anti-Flag antibodies ( H ). WT and 3311m mutant infectious clones were used to transfect WRD cells in which CPSF5 was knocked down, then the cells were subjected to IP with anti-CPSF5 antibodies. qRT-PCR was performed to quantify MVC RNA ( I ). IgG was used as a negative control. Unpaired student’s t -tests were performed, and the data are presented as the means ± SEMs ( n = 3). ***P ≤ 0.001.

Article Snippet: Indicated proteins were detected using primary mouse monoclonal antibodies against beta-actin (sc47778, Santa Cruz Biotechnology, Dallas, TX, USA) and GAPDH (Cat. No. 60004-1-lg; Proteintech); rabbit polyclonal antibodies against CPSF5 (A4482, ABclonal), NAT10 (13365-1-AP, Proteintech), and Flag (F1804-1 MG, Sigma–Aldrich); anti-HA (66006-1-Ig, Proteintech) and anti-Histone H3 antibodies (Cat. No. GTX122148; GeneTex); and three rabbit polyclonal antibodies against MVC NP1, NS1, and VP2 that were previously generated [ ].

Techniques: Purification, Western Blot, Negative Control, Southern Blot, Knockdown, Infection, Expressing, Transfection, Mutagenesis, Control, Quantitative RT-PCR, Modification, Residue, Binding Assay, Clone Assay

Journal: Nucleic Acids Research

Article Title: N4-acetylcytidine coordinates with NP1 and CPSF5 to facilitate alternative RNA processing during the replication of minute virus of canines

doi: 10.1093/nar/gkaf229

Figure Lengend Snippet: MS data of NP1

Techniques:

Journal: Nucleic Acids Research

Article Title: N4-acetylcytidine coordinates with NP1 and CPSF5 to facilitate alternative RNA processing during the replication of minute virus of canines

doi: 10.1093/nar/gkaf229

Figure Lengend Snippet: CPSF5 mediated MVC RNA splicing is dependent on NP1 and ac4C modification at residue 3311. ( A , B ) The effect on CPSF5 binding to MVC RNA by increasing NP1 expression using formaldehyde-RIP-qRT-PCR. Vector or HA-NP1 and infectious clone WT or 3311m were co-transfected in WRD cells. Cell lysates cross-linked by formaldehyde were subjected to IP with IgG, anti-CPSF5 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). *** P ≤ 0.001, ns: not significant. ( C , D ) The effect on NP1 binding to MVC RNA by increasing CPSF5 expression using formaldehyde-RIP-qRT-PCR. Vector or Flag-CPSF5 and infectious clone WT or mutant 3311m were co-transfected in WRD cells. Cell lysates were subjected to IP with IgG, anti-NP1 antibodies. qRT-PCR was performed to quantify the targeted MVC RNA. IgG was used as a negative control. Unpaired Student’s t -tests were performed, and the data are presented as means ± standard errors of the means ( n = 3). ns: not significant. ( E ) Characterization of the cleavage and polyadenylation sites and downstream elements (DSEs) in (pA)p is shown at the top. The bottom panel shows a schematic representation of the RNA pulldown assay. The DNA template for T7 transcription consisted of three repeats of modified DSE. ( F , G ) Biotinylated RNA affinities of NP1 and CPSF5. HA-tagged NP1 or the vector was transiently expressed in WRD cells, and cell lysates were used to bind biotinylated RNA with or without the ac4C modification. In addition, purified NP1 or CPSF5 protein was mixed with RNA oligonucleotides with or without ac4C-modified RNA and analyzed via western blotting. ( H , I ) The binding analysis of CPSF5 or NP1 to oligo-RNA with or without the ac4C modification in the presence or absence of the NP1 ( H ) or CPSF5 ( I ) protein. ( J , K ) The binding analysis of NP1 and CPSF5 to ac4C-modified RNA. Equal amounts of purified CPSF5 ( J ) or NP1 ( K ) protein and increasing amounts of NP1 ( J ) or CPSF5 ( K ) protein were incubated with ac4C-modified RNA and analyzed via western blotting. Blots were detected using anti-CPSF5 or anti-NP1 antibodies.

Techniques: Modification, Residue, Binding Assay, Expressing, Quantitative RT-PCR, Plasmid Preparation, Transfection, Negative Control, Mutagenesis, Purification, Western Blot, Incubation

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