Structured Review

Promega rutp
Rutp, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rutp/product/Promega
Average 90 stars, based on 14 article reviews
Price from $9.99 to $1999.99
rutp - by Bioz Stars, 2020-08
90/100 stars

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Amplification:

Article Title: Pentamidine inhibits catalytic activity of group I intron Ca.LSU by altering RNA folding
Article Snippet: .. The PCR amplified DNA fragment containing the SP6 promoter at the 5′ end of the sense strand of a fragment of the rRNA gene containing Ca.LSU was transcribed in vitro in the presence of 500 µM rATP, rCTP and rGTP, 200 µM rUTP (Promega) and 20 µCi [α-32 P]UTP (3000 Ci/mmol; NEN-DuPont), as previously described ( ). ..

In Vitro:

Article Title: DXO/Rai1 enzymes remove 5′-end FAD and dephospho-CoA caps on RNAs
Article Snippet: .. In vitro transcription was carried out at 37°C for 90 min in 50 μl of the reaction mixture containing 400 ng PCR-generated DNA template, 10 μl of 5 × transcription buffer, 5 μl of 10 mM DTT, 5 μl of 10 μg/μl BSA, 5 μl of 5 mM rCTP, 5 μl of 5 mM rUTP, 20 μCi [α-32P]GTP, 2 μl of T7 RNA polymerase (Promega), and 1 μμl of 40 U/μl RNasin Ribonuclease Inhibitor (Promega). .. Reactions were incubated for 90 min at 37°C and then treated with 3 units of DNase RQ1 (Promega) for 20 min at 37°C.

Article Title: Pentamidine inhibits catalytic activity of group I intron Ca.LSU by altering RNA folding
Article Snippet: .. The PCR amplified DNA fragment containing the SP6 promoter at the 5′ end of the sense strand of a fragment of the rRNA gene containing Ca.LSU was transcribed in vitro in the presence of 500 µM rATP, rCTP and rGTP, 200 µM rUTP (Promega) and 20 µCi [α-32 P]UTP (3000 Ci/mmol; NEN-DuPont), as previously described ( ). ..

Synthesized:

Article Title: Chemokine Gene Expression in Astrocytes of Borna Disease Virus-Infected Rats and Mice in the Absence of Inflammation
Article Snippet: .. The radiolabeled antisense RPA probe set was synthesized in a volume of 20 μl containing 100 μCi of [α-32 P]UTP (3,000 Ci/mmol); dithiothreitol (200 nmol); transcription buffer (Promega); 1 μl of template DNA; rUTP (61 pmol); rGTP, rATP, and rCTP (2.75 nmol each); RNase inhibitor (28 U; Pharmacia); and T7 polymerase (20 U; Promega). .. After 1 h of incubation at 37°C, the template DNAs were digested by treatment with DNase I (4 U; Ambion) for 30 min at 37°C.

Generated:

Article Title: RNA Directed Modulation of Phenotypic Plasticity in Human Cells
Article Snippet: .. Generating Biotin Labeled RNA Transcripts- Biotin labeled RNA transcripts of Retro-EFI2S2 and GFP were generated by adding the following components into an 8 well PCR strip: (2.5μg of DNA template, 2μl T7 Enzyme mix (Sigma), 1ul rATP, rGTP, rCTP, rUTP (Promega), 1μl Biotin-14-CTP (Life Technologies), 2.5μl 1M DTT (Sigma) and 1μl RNase Inhibitor (Life Technologies)(as described in [ ]). .. The reaction volume was made up to 50μl with RNase free water (QIAGEN).

Labeling:

Article Title: RNA Directed Modulation of Phenotypic Plasticity in Human Cells
Article Snippet: .. Generating Biotin Labeled RNA Transcripts- Biotin labeled RNA transcripts of Retro-EFI2S2 and GFP were generated by adding the following components into an 8 well PCR strip: (2.5μg of DNA template, 2μl T7 Enzyme mix (Sigma), 1ul rATP, rGTP, rCTP, rUTP (Promega), 1μl Biotin-14-CTP (Life Technologies), 2.5μl 1M DTT (Sigma) and 1μl RNase Inhibitor (Life Technologies)(as described in [ ]). .. The reaction volume was made up to 50μl with RNase free water (QIAGEN).

Incubation:

Article Title: I-PfoP3I: A Novel Nicking HNH Homing Endonuclease Encoded in the Group I Intron of the DNA Polymerase Gene in Phormidium foveolarum Phage Pf-WMP3
Article Snippet: .. The XhoI linearized pETP3DNAP[+int] (1 µg) was incubated with 40 mM Tris-HCl (pH 7.9), 10 mM NaCl, 6 mM MgCl2 , 2 mM spermidine, 10 mM DTT, 40 u Recombinant RNasin Ribonuclease Inhibitor, 0.5 mM each of rATP, rGTP, rCTP and rUTP, 20 u T7 RNA Polymerase in a total volume of 20 µl at 37°C for 1 h. Template DNA was digested with RNase-free DNaseI (Promega). .. Reverse transcription was performed using sequence-specific primer P3r2 and M-MLV Reverse Transcriptase (Promega) according to the manufacturer’s recommendations.

Article Title: Long-Term Follow-Up of Chimpanzees Inoculated with the First Infectious Clone for Hepatitis C Virus
Article Snippet: .. The 100-μl reaction mixture, containing 20 μl of 5× T7 buffer; 7.5 mM (each) rATP, rCTP, rGTP, and rUTP; 2 μg of DNA; and 10 μl of enzyme mixture was incubated at 37°C for 2 h and then treated with 10 U of RNase-free DNase (Promega) for 30 min at 37°C. ..

Stripping Membranes:

Article Title: RNA Directed Modulation of Phenotypic Plasticity in Human Cells
Article Snippet: .. Generating Biotin Labeled RNA Transcripts- Biotin labeled RNA transcripts of Retro-EFI2S2 and GFP were generated by adding the following components into an 8 well PCR strip: (2.5μg of DNA template, 2μl T7 Enzyme mix (Sigma), 1ul rATP, rGTP, rCTP, rUTP (Promega), 1μl Biotin-14-CTP (Life Technologies), 2.5μl 1M DTT (Sigma) and 1μl RNase Inhibitor (Life Technologies)(as described in [ ]). .. The reaction volume was made up to 50μl with RNase free water (QIAGEN).

Polymerase Chain Reaction:

Article Title: RNA Directed Modulation of Phenotypic Plasticity in Human Cells
Article Snippet: .. Generating Biotin Labeled RNA Transcripts- Biotin labeled RNA transcripts of Retro-EFI2S2 and GFP were generated by adding the following components into an 8 well PCR strip: (2.5μg of DNA template, 2μl T7 Enzyme mix (Sigma), 1ul rATP, rGTP, rCTP, rUTP (Promega), 1μl Biotin-14-CTP (Life Technologies), 2.5μl 1M DTT (Sigma) and 1μl RNase Inhibitor (Life Technologies)(as described in [ ]). .. The reaction volume was made up to 50μl with RNase free water (QIAGEN).

Article Title: DXO/Rai1 enzymes remove 5′-end FAD and dephospho-CoA caps on RNAs
Article Snippet: .. In vitro transcription was carried out at 37°C for 90 min in 50 μl of the reaction mixture containing 400 ng PCR-generated DNA template, 10 μl of 5 × transcription buffer, 5 μl of 10 mM DTT, 5 μl of 10 μg/μl BSA, 5 μl of 5 mM rCTP, 5 μl of 5 mM rUTP, 20 μCi [α-32P]GTP, 2 μl of T7 RNA polymerase (Promega), and 1 μμl of 40 U/μl RNasin Ribonuclease Inhibitor (Promega). .. Reactions were incubated for 90 min at 37°C and then treated with 3 units of DNase RQ1 (Promega) for 20 min at 37°C.

Article Title: Pentamidine inhibits catalytic activity of group I intron Ca.LSU by altering RNA folding
Article Snippet: .. The PCR amplified DNA fragment containing the SP6 promoter at the 5′ end of the sense strand of a fragment of the rRNA gene containing Ca.LSU was transcribed in vitro in the presence of 500 µM rATP, rCTP and rGTP, 200 µM rUTP (Promega) and 20 µCi [α-32 P]UTP (3000 Ci/mmol; NEN-DuPont), as previously described ( ). ..

Recombinant:

Article Title: I-PfoP3I: A Novel Nicking HNH Homing Endonuclease Encoded in the Group I Intron of the DNA Polymerase Gene in Phormidium foveolarum Phage Pf-WMP3
Article Snippet: .. The XhoI linearized pETP3DNAP[+int] (1 µg) was incubated with 40 mM Tris-HCl (pH 7.9), 10 mM NaCl, 6 mM MgCl2 , 2 mM spermidine, 10 mM DTT, 40 u Recombinant RNasin Ribonuclease Inhibitor, 0.5 mM each of rATP, rGTP, rCTP and rUTP, 20 u T7 RNA Polymerase in a total volume of 20 µl at 37°C for 1 h. Template DNA was digested with RNase-free DNaseI (Promega). .. Reverse transcription was performed using sequence-specific primer P3r2 and M-MLV Reverse Transcriptase (Promega) according to the manufacturer’s recommendations.

Recombinase Polymerase Amplification:

Article Title: Chemokine Gene Expression in Astrocytes of Borna Disease Virus-Infected Rats and Mice in the Absence of Inflammation
Article Snippet: .. The radiolabeled antisense RPA probe set was synthesized in a volume of 20 μl containing 100 μCi of [α-32 P]UTP (3,000 Ci/mmol); dithiothreitol (200 nmol); transcription buffer (Promega); 1 μl of template DNA; rUTP (61 pmol); rGTP, rATP, and rCTP (2.75 nmol each); RNase inhibitor (28 U; Pharmacia); and T7 polymerase (20 U; Promega). .. After 1 h of incubation at 37°C, the template DNAs were digested by treatment with DNase I (4 U; Ambion) for 30 min at 37°C.

Plasmid Preparation:

Article Title: TALEN/CRISPR-mediated engineering of a promoterless anti-viral RNAi hairpin into an endogenous miRNA locus
Article Snippet: .. For HCV transcripts and use with SP6 polymerase, the mix contained 80 mM HEPES (pH 7.5), 16 mM MgCl2 , 2 mM spermidine, 40 mM DTT, 3.125 mM each of rATP, rCTP and rUTP, 1.5625 mM of rGTP, 1 mM m7 G(5′)ppp(5′)G RNA cap structure analog, 1 U/μl of RNasin, 0.1 μg/μl of plasmid DNA and 0.6 U/μl of SP6 RNA polymerase (Promega). .. After a 2 h incubation at 37°C (HCV) or 40°C (Dengue virus), 0.3 U of T7 RNA polymerase or 0.4 U of SP6 RNA polymerase, respectively, were added per μl of reaction mixture, and the reaction mixture was incubated overnight.

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  • 85
    Promega alpha 32p rutp
    Prep1 inhibits translation of the luciferase- Hoxb4 mRNA and is able to bind the 3′-UTR region of Hoxb4 mRNA. (A) Luc-Hoxb4 3′UTR mRNA was translated in vitro in a rabbit reticulocytes system with the additions indicated at the bottom (Prep1 or 4EHP previously in vitro synthesized under T7 promoter). Luciferase activity is expressed in percent of the control; the 100% value is the level of luciferase translated in the absence of any added protein (column 6). Addition of in vitro translated Prep1 inhibits Luc-Hoxb4 3′UTR mRNA translation (columns 1 and 4) while no further effect is observed when the in vitro translated 4EHP protein is added to the reaction with Prep1 (column 1) or alone (column 3). Less inhibition is obtained with the in vitro translated Prep1 4EHP-binding (Y-LL) mutant (Prep1-Mut) (columns 2 and 5). N-values are 5. (B) This control shows that the differences observed in (A) are not due to interference with the in vitro transcription of the luciferase- Hoxb4 3′UTR mRNA. RT-PCR analysis of Luc-Hoxb4 3′UTR mRNA present in samples 1–6 (A). Each reaction was amplified for 25 and 30 cycles. Notice the absence of amplification in the RT(-), indicating that the plasmid used for Luc-Hoxb4 3′UTR transcription had been completely digested by the DNAse treatment. (C) Prep1 does not inhibit translation of Luc-Cdx2 3′UTR mRNA, independently of the presence of 4EHP. Thus the inhibitory effect appears to be dictated by the presence of the Hoxb4 3′UTR. N-values are 3. (D) Prep1 inhibits in vitro translation of Luc-Hoxb4 3′UTR mRNA in a dose-dependent manner. Compare non diluted Prep1 (column 1) with dilutions 1/2 and 1/5 (columns 2 and 3). N-values are 4. (E) Anti-4EHP antibodies prevent the inhibitory action of Prep1. Inhibition of Luc-Hoxb4 3′UTR mRNA translation by Prep1 was reversed when 2 µg of anti-4EHP (but not an unrelated) antibody was added to the reaction. N-values are 3. (F) RNA-EMSA showing specific Prep1 binding to Hoxb4 3′UTR. First lane shows the <t>[alpha-32P]</t> <t>rUTP-labelled</t> probe. Second lane shows the shift induced by the addition of Prep1 to the reaction (arrow). Third lane shows the induction of a super-shift by the Prep1 antibody (arrowhead). Lane 4 shows that the effect of the antibody is specific since an anti-Pbx antibody has no effect. Same experiment using antisense probe is shown in lanes 5–8.
    Alpha 32p Rutp, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alpha 32p rutp/product/Promega
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alpha 32p rutp - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

    90
    Promega rutp
    Prep1 inhibits translation of the luciferase- Hoxb4 mRNA and is able to bind the 3′-UTR region of Hoxb4 mRNA. (A) Luc-Hoxb4 3′UTR mRNA was translated in vitro in a rabbit reticulocytes system with the additions indicated at the bottom (Prep1 or 4EHP previously in vitro synthesized under T7 promoter). Luciferase activity is expressed in percent of the control; the 100% value is the level of luciferase translated in the absence of any added protein (column 6). Addition of in vitro translated Prep1 inhibits Luc-Hoxb4 3′UTR mRNA translation (columns 1 and 4) while no further effect is observed when the in vitro translated 4EHP protein is added to the reaction with Prep1 (column 1) or alone (column 3). Less inhibition is obtained with the in vitro translated Prep1 4EHP-binding (Y-LL) mutant (Prep1-Mut) (columns 2 and 5). N-values are 5. (B) This control shows that the differences observed in (A) are not due to interference with the in vitro transcription of the luciferase- Hoxb4 3′UTR mRNA. RT-PCR analysis of Luc-Hoxb4 3′UTR mRNA present in samples 1–6 (A). Each reaction was amplified for 25 and 30 cycles. Notice the absence of amplification in the RT(-), indicating that the plasmid used for Luc-Hoxb4 3′UTR transcription had been completely digested by the DNAse treatment. (C) Prep1 does not inhibit translation of Luc-Cdx2 3′UTR mRNA, independently of the presence of 4EHP. Thus the inhibitory effect appears to be dictated by the presence of the Hoxb4 3′UTR. N-values are 3. (D) Prep1 inhibits in vitro translation of Luc-Hoxb4 3′UTR mRNA in a dose-dependent manner. Compare non diluted Prep1 (column 1) with dilutions 1/2 and 1/5 (columns 2 and 3). N-values are 4. (E) Anti-4EHP antibodies prevent the inhibitory action of Prep1. Inhibition of Luc-Hoxb4 3′UTR mRNA translation by Prep1 was reversed when 2 µg of anti-4EHP (but not an unrelated) antibody was added to the reaction. N-values are 3. (F) RNA-EMSA showing specific Prep1 binding to Hoxb4 3′UTR. First lane shows the <t>[alpha-32P]</t> <t>rUTP-labelled</t> probe. Second lane shows the shift induced by the addition of Prep1 to the reaction (arrow). Third lane shows the induction of a super-shift by the Prep1 antibody (arrowhead). Lane 4 shows that the effect of the antibody is specific since an anti-Pbx antibody has no effect. Same experiment using antisense probe is shown in lanes 5–8.
    Rutp, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rutp/product/Promega
    Average 90 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    rutp - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    80
    Promega p rutp labeled antisense riboprobes
    Prep1 inhibits translation of the luciferase- Hoxb4 mRNA and is able to bind the 3′-UTR region of Hoxb4 mRNA. (A) Luc-Hoxb4 3′UTR mRNA was translated in vitro in a rabbit reticulocytes system with the additions indicated at the bottom (Prep1 or 4EHP previously in vitro synthesized under T7 promoter). Luciferase activity is expressed in percent of the control; the 100% value is the level of luciferase translated in the absence of any added protein (column 6). Addition of in vitro translated Prep1 inhibits Luc-Hoxb4 3′UTR mRNA translation (columns 1 and 4) while no further effect is observed when the in vitro translated 4EHP protein is added to the reaction with Prep1 (column 1) or alone (column 3). Less inhibition is obtained with the in vitro translated Prep1 4EHP-binding (Y-LL) mutant (Prep1-Mut) (columns 2 and 5). N-values are 5. (B) This control shows that the differences observed in (A) are not due to interference with the in vitro transcription of the luciferase- Hoxb4 3′UTR mRNA. RT-PCR analysis of Luc-Hoxb4 3′UTR mRNA present in samples 1–6 (A). Each reaction was amplified for 25 and 30 cycles. Notice the absence of amplification in the RT(-), indicating that the plasmid used for Luc-Hoxb4 3′UTR transcription had been completely digested by the DNAse treatment. (C) Prep1 does not inhibit translation of Luc-Cdx2 3′UTR mRNA, independently of the presence of 4EHP. Thus the inhibitory effect appears to be dictated by the presence of the Hoxb4 3′UTR. N-values are 3. (D) Prep1 inhibits in vitro translation of Luc-Hoxb4 3′UTR mRNA in a dose-dependent manner. Compare non diluted Prep1 (column 1) with dilutions 1/2 and 1/5 (columns 2 and 3). N-values are 4. (E) Anti-4EHP antibodies prevent the inhibitory action of Prep1. Inhibition of Luc-Hoxb4 3′UTR mRNA translation by Prep1 was reversed when 2 µg of anti-4EHP (but not an unrelated) antibody was added to the reaction. N-values are 3. (F) RNA-EMSA showing specific Prep1 binding to Hoxb4 3′UTR. First lane shows the <t>[alpha-32P]</t> <t>rUTP-labelled</t> probe. Second lane shows the shift induced by the addition of Prep1 to the reaction (arrow). Third lane shows the induction of a super-shift by the Prep1 antibody (arrowhead). Lane 4 shows that the effect of the antibody is specific since an anti-Pbx antibody has no effect. Same experiment using antisense probe is shown in lanes 5–8.
    P Rutp Labeled Antisense Riboprobes, supplied by Promega, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p rutp labeled antisense riboprobes/product/Promega
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p rutp labeled antisense riboprobes - by Bioz Stars, 2020-08
    80/100 stars
      Buy from Supplier

    Image Search Results


    Prep1 inhibits translation of the luciferase- Hoxb4 mRNA and is able to bind the 3′-UTR region of Hoxb4 mRNA. (A) Luc-Hoxb4 3′UTR mRNA was translated in vitro in a rabbit reticulocytes system with the additions indicated at the bottom (Prep1 or 4EHP previously in vitro synthesized under T7 promoter). Luciferase activity is expressed in percent of the control; the 100% value is the level of luciferase translated in the absence of any added protein (column 6). Addition of in vitro translated Prep1 inhibits Luc-Hoxb4 3′UTR mRNA translation (columns 1 and 4) while no further effect is observed when the in vitro translated 4EHP protein is added to the reaction with Prep1 (column 1) or alone (column 3). Less inhibition is obtained with the in vitro translated Prep1 4EHP-binding (Y-LL) mutant (Prep1-Mut) (columns 2 and 5). N-values are 5. (B) This control shows that the differences observed in (A) are not due to interference with the in vitro transcription of the luciferase- Hoxb4 3′UTR mRNA. RT-PCR analysis of Luc-Hoxb4 3′UTR mRNA present in samples 1–6 (A). Each reaction was amplified for 25 and 30 cycles. Notice the absence of amplification in the RT(-), indicating that the plasmid used for Luc-Hoxb4 3′UTR transcription had been completely digested by the DNAse treatment. (C) Prep1 does not inhibit translation of Luc-Cdx2 3′UTR mRNA, independently of the presence of 4EHP. Thus the inhibitory effect appears to be dictated by the presence of the Hoxb4 3′UTR. N-values are 3. (D) Prep1 inhibits in vitro translation of Luc-Hoxb4 3′UTR mRNA in a dose-dependent manner. Compare non diluted Prep1 (column 1) with dilutions 1/2 and 1/5 (columns 2 and 3). N-values are 4. (E) Anti-4EHP antibodies prevent the inhibitory action of Prep1. Inhibition of Luc-Hoxb4 3′UTR mRNA translation by Prep1 was reversed when 2 µg of anti-4EHP (but not an unrelated) antibody was added to the reaction. N-values are 3. (F) RNA-EMSA showing specific Prep1 binding to Hoxb4 3′UTR. First lane shows the [alpha-32P] rUTP-labelled probe. Second lane shows the shift induced by the addition of Prep1 to the reaction (arrow). Third lane shows the induction of a super-shift by the Prep1 antibody (arrowhead). Lane 4 shows that the effect of the antibody is specific since an anti-Pbx antibody has no effect. Same experiment using antisense probe is shown in lanes 5–8.

    Journal: PLoS ONE

    Article Title: Cytoplasmic Prep1 Interacts with 4EHP Inhibiting Hoxb4 Translation

    doi: 10.1371/journal.pone.0005213

    Figure Lengend Snippet: Prep1 inhibits translation of the luciferase- Hoxb4 mRNA and is able to bind the 3′-UTR region of Hoxb4 mRNA. (A) Luc-Hoxb4 3′UTR mRNA was translated in vitro in a rabbit reticulocytes system with the additions indicated at the bottom (Prep1 or 4EHP previously in vitro synthesized under T7 promoter). Luciferase activity is expressed in percent of the control; the 100% value is the level of luciferase translated in the absence of any added protein (column 6). Addition of in vitro translated Prep1 inhibits Luc-Hoxb4 3′UTR mRNA translation (columns 1 and 4) while no further effect is observed when the in vitro translated 4EHP protein is added to the reaction with Prep1 (column 1) or alone (column 3). Less inhibition is obtained with the in vitro translated Prep1 4EHP-binding (Y-LL) mutant (Prep1-Mut) (columns 2 and 5). N-values are 5. (B) This control shows that the differences observed in (A) are not due to interference with the in vitro transcription of the luciferase- Hoxb4 3′UTR mRNA. RT-PCR analysis of Luc-Hoxb4 3′UTR mRNA present in samples 1–6 (A). Each reaction was amplified for 25 and 30 cycles. Notice the absence of amplification in the RT(-), indicating that the plasmid used for Luc-Hoxb4 3′UTR transcription had been completely digested by the DNAse treatment. (C) Prep1 does not inhibit translation of Luc-Cdx2 3′UTR mRNA, independently of the presence of 4EHP. Thus the inhibitory effect appears to be dictated by the presence of the Hoxb4 3′UTR. N-values are 3. (D) Prep1 inhibits in vitro translation of Luc-Hoxb4 3′UTR mRNA in a dose-dependent manner. Compare non diluted Prep1 (column 1) with dilutions 1/2 and 1/5 (columns 2 and 3). N-values are 4. (E) Anti-4EHP antibodies prevent the inhibitory action of Prep1. Inhibition of Luc-Hoxb4 3′UTR mRNA translation by Prep1 was reversed when 2 µg of anti-4EHP (but not an unrelated) antibody was added to the reaction. N-values are 3. (F) RNA-EMSA showing specific Prep1 binding to Hoxb4 3′UTR. First lane shows the [alpha-32P] rUTP-labelled probe. Second lane shows the shift induced by the addition of Prep1 to the reaction (arrow). Third lane shows the induction of a super-shift by the Prep1 antibody (arrowhead). Lane 4 shows that the effect of the antibody is specific since an anti-Pbx antibody has no effect. Same experiment using antisense probe is shown in lanes 5–8.

    Article Snippet: REMSA The probe for REMSA was prepared and labeled by the in vitro transcription of the cloned DNA fragment of Hoxb4 3′UTR using [alpha-32P]rUTP and Riboprobe® Combination System (Promega, Madison, WI).

    Techniques: Luciferase, In Vitro, Synthesized, Activity Assay, Inhibition, Binding Assay, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Amplification, Plasmid Preparation