Recombinant Rat MATR3 Protein

• Cat.No.: MATR3-3598R
• Product Overview: Recombinant Rat MATR3 full length or partial length protein was expressed.

Specification

• Species: Rat
• Source: Mammalian Cells
• Tag: His
• Form: Liquid or lyophilized powder
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: >80%
• Notes: This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
• Storage: Store it at +4 ºC for short term. For long term storage, store it at -20 ºC~-80 ºC.
• Storage Buffer: PBS buffer

Gene Information

• Gene Name: Matr3 matrin 3 [ Rattus norvegicus ]
• Official Symbol: MATR3
• Gene ID: 29150
• mRNA Refseq: NM_019149.3
• Protein Refseq: NP_062022.2
• UniProt ID: P43244

Citation

Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3

Journal: eLife    PubMed ID: 28580901    Data: 2017/6/5

Authors: Ritu Chaudhary, Berkley Gryder, Joaquín M Espinosa

Article Snippet:Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.. To determine the direct binding of PINCR to recombinant Matrin 3 (rMatrin 3), 200 ng of in vitro transcribed Bi- PINCR or Bi- LUC RNA was incubated with 500 ng recombinant Matrin 3 protein (Creative BioMart, Catalog # MATR3-15H) in 1X EMSA buffer (25 mM Tris-HCl pH 7.5, 150 mM KCl, 0.1% Triton-X-100, 100 μg/ml BSA, 2 mM DTT and 5% glycerol) at room temperature for 2 hr.. RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.

( A ) Peptide spectrum matches (PSMs) corresponding to Matrin 3 in the Bi- LUC and Bi- PINCR pulldowns from mass spectrometry analysis. ( B, C ) Streptavidin pulldowns followed by immunoblotting was performed following incubation of Bi- LUC and Bi- PINCR RNA with DOXO-treated HCT116 nuclear extracts ( B ) or recombinant Matrin 3 (rMatrin 3) ( C ). ( D ) Specific enrichment of PINCR in the Matrin 3 IPs was assessed by qRT-PCR from 24 hr 5-FU-treated formaldehyde cross-linked HCT116 cells. p21 mRNA was used as negative control. ( E ) PINCR -WT cells were transfected with CTL or two independent Matrin 3 siRNAs (I and II) for 48 hr and Matrin 3 knockdown was measured by immunoblotting. ( F ) PINCR -WT and PINCR -KO cells were transfected with CTL or Matrin 3 siRNAs and after 48 hr the cells were untreated or treated with 5-FU for 48 hr. The effect on the sub-G1 population was assessed by PI staining followed by FACS. ( G ) PINCR -WT and PINCR -KO cells were transfected with CTL or Matrin 3 siRNAs for 48 hr; transfected cells were left untreated or treated with 5-FU for 24 hr and qRT-PCR was performed. Error bars in D, F and G represent SD from three independent experiments. *p<0.05; # p <0.01; **p<0.005. DOI: http://dx.doi.org/10.7554/eLife.23244.038 10.7554/eLife.23244.039 Figure 5—source data 1. Matrin 3 immunoblot for . DOI: http://dx.doi.org/10.7554/eLife.23244.039

Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3

Journal: eLife    PubMed ID: 28580901    Data: 2017/6/5

Authors: Ritu Chaudhary, Berkley Gryder, Joaquín M Espinosa

Article Snippet:Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.. To determine the direct binding of PINCR to recombinant Matrin 3 (rMatrin 3), 200 ng of in vitro transcribed Bi- PINCR or Bi- LUC RNA was incubated with 500 ng recombinant Matrin 3 protein (Creative BioMart, Catalog # MATR3-15H) in 1X EMSA buffer (25 mM Tris-HCl pH 7.5, 150 mM KCl, 0.1% Triton-X-100, 100 μg/ml BSA, 2 mM DTT and 5% glycerol) at room temperature for 2 hr.. RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.

( A, B ) Putative Matrin 3 binding motif in PINCR RNA. ( A ) ‘N’ represents the number of times the motif appears in the PINCR RNA. ‘ES’ represents the enrichment score calculated as shown in ‘ B ’. ( C ) HCT116 cells were reverse transfected with a control siRNA (siCTL) or siRNAs targeting Matrin 3 (siMatrin 3-I and siMatrin 3-II) for 48 hr and the extent of Matrin 3 knockdown was measured by qRT-PCR for Matrin 3 normalized to GADPH. Error bars represent SD from three independent experiments. ##p<0.001. DOI: http://dx.doi.org/10.7554/eLife.23244.040

PINCR -WT and PINCR -KO HCT116 cells were reverse transfected with a control siRNA (siCTL) or siRNAs targeting Matrin 3 (siMatrin 3-I and siMatrin 3-II) for 48 hr. Transfected cells were left untreated or treated with DOXO or 5-FU for 48 hr. The effect on G1 arrest and apoptosis was examined by PI-staining and FACS analysis. Shown are the results from three independent experiments after 5-FU treatment ( A ) or DOXO treatment ( B, C ). Error bars represent SD from three independent experiments. #p<0.01, ##p<0.001. DOI: http://dx.doi.org/10.7554/eLife.23244.041

Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3

Journal: eLife    PubMed ID: 28580901    Data: 2017/6/5

Authors: Ritu Chaudhary, Berkley Gryder, Joaquín M Espinosa

Article Snippet:Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.Matrin 3 bound RNAs were isolated by phenol-chloroform extraction (Ambion) followed by ethanol precipitation and qRT-PCR was used to determine the enrichment of p21 (negative control) and PINCR in the Matrin 3 IPs.. To determine the direct binding of PINCR to recombinant Matrin 3 (rMatrin 3), 200 ng of in vitro transcribed Bi- PINCR or Bi- LUC RNA was incubated with 500 ng recombinant Matrin 3 protein (Creative BioMart, Catalog # MATR3-15H) in 1X EMSA buffer (25 mM Tris-HCl pH 7.5, 150 mM KCl, 0.1% Triton-X-100, 100 μg/ml BSA, 2 mM DTT and 5% glycerol) at room temperature for 2 hr.. RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.RNA–protein complex was immunoprecipitated at room temperature for 2 hr, by using Dynabeads M-280 Streptavidin.

( A ) Peptide spectrum matches (PSMs) corresponding to Matrin 3 in the Bi- LUC and Bi- PINCR pulldowns from mass spectrometry analysis. ( B, C ) Streptavidin pulldowns followed by immunoblotting was performed following incubation of Bi- LUC and Bi- PINCR RNA with DOXO-treated HCT116 nuclear extracts ( B ) or recombinant Matrin 3 (rMatrin 3) ( C ). ( D ) Specific enrichment of PINCR in the Matrin 3 IPs was assessed by qRT-PCR from 24 hr 5-FU-treated formaldehyde cross-linked HCT116 cells. p21 mRNA was used as negative control. ( E ) PINCR -WT cells were transfected with CTL or two independent Matrin 3 siRNAs (I and II) for 48 hr and Matrin 3 knockdown was measured by immunoblotting. ( F ) PINCR -WT and PINCR -KO cells were transfected with CTL or Matrin 3 siRNAs and after 48 hr the cells were untreated or treated with 5-FU for 48 hr. The effect on the sub-G1 population was assessed by PI staining followed by FACS. ( G ) PINCR -WT and PINCR -KO cells were transfected with CTL or Matrin 3 siRNAs for 48 hr; transfected cells were left untreated or treated with 5-FU for 24 hr and qRT-PCR was performed. Error bars in D, F and G represent SD from three independent experiments. *p<0.05; # p <0.01; **p<0.005. DOI: http://dx.doi.org/10.7554/eLife.23244.038 10.7554/eLife.23244.039 Figure 5—source data 1. Matrin 3 immunoblot for . DOI: http://dx.doi.org/10.7554/eLife.23244.039

( A, B ) Putative Matrin 3 binding motif in PINCR RNA. ( A ) ‘N’ represents the number of times the motif appears in the PINCR RNA. ‘ES’ represents the enrichment score calculated as shown in ‘ B ’. ( C ) HCT116 cells were reverse transfected with a control siRNA (siCTL) or siRNAs targeting Matrin 3 (siMatrin 3-I and siMatrin 3-II) for 48 hr and the extent of Matrin 3 knockdown was measured by qRT-PCR for Matrin 3 normalized to GADPH. Error bars represent SD from three independent experiments. ##p<0.001. DOI: http://dx.doi.org/10.7554/eLife.23244.040

PINCR -WT and PINCR -KO HCT116 cells were reverse transfected with a control siRNA (siCTL) or siRNAs targeting Matrin 3 (siMatrin 3-I and siMatrin 3-II) for 48 hr. Transfected cells were left untreated or treated with DOXO or 5-FU for 48 hr. The effect on G1 arrest and apoptosis was examined by PI-staining and FACS analysis. Shown are the results from three independent experiments after 5-FU treatment ( A ) or DOXO treatment ( B, C ). Error bars represent SD from three independent experiments. #p<0.01, ##p<0.001. DOI: http://dx.doi.org/10.7554/eLife.23244.041