superase ln  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 88

    Structured Review

    Thermo Fisher superase ln
    Superase Ln, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superase ln/product/Thermo Fisher
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    superase ln - by Bioz Stars, 2020-09
    88/100 stars

    Images

    Related Articles

    Random Hexamer Labeling:

    Article Title: Compartmentalized microbial composition, oxygen gradients and nitrogen fixation in the gut of Odontotaenius disjunctus
    Article Snippet: .. Total RNA, 300 ng of random hexamer primers and 20 nmol of each dNTP were combined to a total volume of 11 μl and the mixture incubated at 65 °C for 5 min. Then, 5 μl of 5 × first-strand buffer was added to the mixture, together with 20 U of SUPERase-ln (Invitrogen, Grand Island, NY, USA) and incubated at 25 °C for 2 min. After incubating, 200 U of SuperScript III reverse transcriptase (Invitrogen) was added to the mixture and incubated at 50 °C for 50 min. .. The reaction was inactivated at 70 °C for 10 min. cDNA was then used as template for quantitiative PCR (qPCR) of the nifH (nitrogenase gene) and rpoB (RNA polymerase β-subunit) genes using the primers PolF and PolR, and 1689F and 2041R ( ; ). qPCR conditions were optimized using a temperature gradient.

    Article Title: Gut microbiota mediate caffeine detoxification in the primary insect pest of coffee
    Article Snippet: .. Total RNA, 300 ng of random hexamer primers and 20 nmol of each dNTP were combined to a total volume of 11 μl and the mixture incubated at 65 °C for 5 min. Then, 5 μl of 5 × first strand buffer was added to the mixture, together with 20 U of SUPERase-ln (Invitrogen), and incubated at 25 °C for 2 min. After incubating, 200 U of SuperScript III reverse transcriptase (Invitrogen) was added to the mixture and incubated at 50 °C for 50 min. .. The reaction was inactivated at 70 °C for 10 min. cDNA was then used as a template for qPCR with the primers CBBcdmF and CBBcdmR.

    Incubation:

    Article Title: Compartmentalized microbial composition, oxygen gradients and nitrogen fixation in the gut of Odontotaenius disjunctus
    Article Snippet: .. Total RNA, 300 ng of random hexamer primers and 20 nmol of each dNTP were combined to a total volume of 11 μl and the mixture incubated at 65 °C for 5 min. Then, 5 μl of 5 × first-strand buffer was added to the mixture, together with 20 U of SUPERase-ln (Invitrogen, Grand Island, NY, USA) and incubated at 25 °C for 2 min. After incubating, 200 U of SuperScript III reverse transcriptase (Invitrogen) was added to the mixture and incubated at 50 °C for 50 min. .. The reaction was inactivated at 70 °C for 10 min. cDNA was then used as template for quantitiative PCR (qPCR) of the nifH (nitrogenase gene) and rpoB (RNA polymerase β-subunit) genes using the primers PolF and PolR, and 1689F and 2041R ( ; ). qPCR conditions were optimized using a temperature gradient.

    Article Title: Gut microbiota mediate caffeine detoxification in the primary insect pest of coffee
    Article Snippet: .. Total RNA, 300 ng of random hexamer primers and 20 nmol of each dNTP were combined to a total volume of 11 μl and the mixture incubated at 65 °C for 5 min. Then, 5 μl of 5 × first strand buffer was added to the mixture, together with 20 U of SUPERase-ln (Invitrogen), and incubated at 25 °C for 2 min. After incubating, 200 U of SuperScript III reverse transcriptase (Invitrogen) was added to the mixture and incubated at 50 °C for 50 min. .. The reaction was inactivated at 70 °C for 10 min. cDNA was then used as a template for qPCR with the primers CBBcdmF and CBBcdmR.

    Synthesized:

    Article Title: Peanut oral immunotherapy transiently expands circulating Ara h 2-specific B cells with a homologous repertoire in unrelated individuals
    Article Snippet: .. Briefly, cDNA was synthesized using Superscript III Reverse Transcriptase (Invitrogen) and five constant region reverse transcriptase primers along with SUPERase-ln (Ambion). .. At the completion, RNAse H (Invitrogen) was used before PCR.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher superase• in rnase inhibitor
    A variety of B lymphocyte lineages from human tonsil are susceptible to infection with BAC16 KSHV. Magnetically sorted total B lymphocytes from four tonsil specimens were infected with KSHV or mock-infected and analyzed by FCM at indicated timepoints for (A) GFP expression and (B) immunophenotypic markers for lineage. In both cases, cells were gated for singlet/viable/CD19+. Memory B cells were further defined as CD38low/IgD-/CD27+, naïve B cells were CD38low/IgD+/CD27-, natural effector (Nat Effector) cells were CD38low/IgD+/CD27+ and germinal center (GC) cells were CD38hi/IgD-. (C) In similar infection experiments with four tonsil specimens, total <t>RNA</t> was extracted at 2, 4 and 6 days post-infection and viral gene transcription was verified in two technical replicates by <t>RT-PCR.</t> Replicate RT negative cDNA reactions for KSHV infected samples at 6 days post-infection were included as a control for DNA contamination and mean NRT Cq values (n = 8) for each target were as follows: 39.44 for LANA, 40.52 for ORF59 and > 40 (not detectable) for K8.1. For a 40-cycle reaction, non-amplifying samples were set to Cq = 41 for the purposes of calculation. The lowest Cq value obtained in a mock infected sample was assigned as the limit of detection for each target, and data points that fall below this threshold are denoted with red shading. Yellow shading highlights values between 1.7 and 3.3 cycles lower than the limit of detection and corresponds to 5–10 fold increases in gene expression. Green shading highlights values more than 3.3 cycles lower than the limit of detection and corresponds to gene expression levels greater than 10-fold above the limit of detection. ANOVA analysis of raw Cq values revealed a statistically significant effect of KSHV infection for all target genes when comparing aggregate trends for mock vs KSHV samples over time: LANA p = 0.0006; K8.1 p = 0.02, ORF59 p
    Superase• In Rnase Inhibitor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superase• in rnase inhibitor/product/Thermo Fisher
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    superase• in rnase inhibitor - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Thermo Fisher cas13b crrna processing buffer
    Biochemical characterization of RNA targeting by <t>Cas13b.</t> A. Thermal melting curves of PbuCas13b complexed with <t>crRNA</t> and target RNA. To the right of the figure legends are cartoons of the substrate used with red lines indicating mismatches (tandem mismatches are shown on the left panel; larger mismatches are shown on the right panel). Curves shifted to the left indicate a destabilized complex. B. The effect of single nucleotide mismatches between target and crRNA spacer on collateral nuclease activity activation in FLUORESCENT COLLATERAL RNA–CLEAVAGE assays. Two different guide sequences (upper, guide 1; lower, guide 2) are shown. C. . D. Diagram of the active site of PbuCas13b in the crystal structure. Citrate (shown in yellow) is bound in the active site and loop 938–951 is shown in the lower left labeled as ‘Loop’.
    Cas13b Crrna Processing Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cas13b crrna processing buffer/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cas13b crrna processing buffer - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    99
    Thermo Fisher superase in rnase inhibitor
    Unique binding motifs in the N-terminus of SLBP interact with FEM1A, FEM1B, and FEM1C. (A) Diagram representing the domain structure of SLBP. The amino acid sequence and substrate receptor binding motifs representing the “degron hotspot” are shown. TAD, translational activation domain; NLS, nuclear localization sequence; RBD, RNA binding domain. (B) FEM1A, FEM1B, and FEM1C interact with amino acids 1–99 of SLBP. C-E Mapping the FEM1A, FEM1B and FEM1C binding regions in SLBP. HEK293T cells were transfected with either empty vector (EV) or FS-tagged SLBP constructs. MLN4924 was added to the cells for 4 hours before collection. Cell lysates were affinity precipitated with anti-STREP resin, and affinity precipitations were probed with the indicated antibodies. (F) The ligase-deficient SLBP(ABCdegron) mutant is unable to bind to CTIF. HEK293T cells were transfected with FLAG-tagged SLBP constructs. Cell lysates were supplemented with <t>SUPERase-In™</t> <t>RNase</t> Inhibitor and immunoprecipitated with anti-FLAG resin. The immunoprecipitations were probed with the indicated antibodies.
    Superase In Rnase Inhibitor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superase in rnase inhibitor/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    superase in rnase inhibitor - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    A variety of B lymphocyte lineages from human tonsil are susceptible to infection with BAC16 KSHV. Magnetically sorted total B lymphocytes from four tonsil specimens were infected with KSHV or mock-infected and analyzed by FCM at indicated timepoints for (A) GFP expression and (B) immunophenotypic markers for lineage. In both cases, cells were gated for singlet/viable/CD19+. Memory B cells were further defined as CD38low/IgD-/CD27+, naïve B cells were CD38low/IgD+/CD27-, natural effector (Nat Effector) cells were CD38low/IgD+/CD27+ and germinal center (GC) cells were CD38hi/IgD-. (C) In similar infection experiments with four tonsil specimens, total RNA was extracted at 2, 4 and 6 days post-infection and viral gene transcription was verified in two technical replicates by RT-PCR. Replicate RT negative cDNA reactions for KSHV infected samples at 6 days post-infection were included as a control for DNA contamination and mean NRT Cq values (n = 8) for each target were as follows: 39.44 for LANA, 40.52 for ORF59 and > 40 (not detectable) for K8.1. For a 40-cycle reaction, non-amplifying samples were set to Cq = 41 for the purposes of calculation. The lowest Cq value obtained in a mock infected sample was assigned as the limit of detection for each target, and data points that fall below this threshold are denoted with red shading. Yellow shading highlights values between 1.7 and 3.3 cycles lower than the limit of detection and corresponds to 5–10 fold increases in gene expression. Green shading highlights values more than 3.3 cycles lower than the limit of detection and corresponds to gene expression levels greater than 10-fold above the limit of detection. ANOVA analysis of raw Cq values revealed a statistically significant effect of KSHV infection for all target genes when comparing aggregate trends for mock vs KSHV samples over time: LANA p = 0.0006; K8.1 p = 0.02, ORF59 p

    Journal: PLoS Pathogens

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes

    doi: 10.1371/journal.ppat.1006967

    Figure Lengend Snippet: A variety of B lymphocyte lineages from human tonsil are susceptible to infection with BAC16 KSHV. Magnetically sorted total B lymphocytes from four tonsil specimens were infected with KSHV or mock-infected and analyzed by FCM at indicated timepoints for (A) GFP expression and (B) immunophenotypic markers for lineage. In both cases, cells were gated for singlet/viable/CD19+. Memory B cells were further defined as CD38low/IgD-/CD27+, naïve B cells were CD38low/IgD+/CD27-, natural effector (Nat Effector) cells were CD38low/IgD+/CD27+ and germinal center (GC) cells were CD38hi/IgD-. (C) In similar infection experiments with four tonsil specimens, total RNA was extracted at 2, 4 and 6 days post-infection and viral gene transcription was verified in two technical replicates by RT-PCR. Replicate RT negative cDNA reactions for KSHV infected samples at 6 days post-infection were included as a control for DNA contamination and mean NRT Cq values (n = 8) for each target were as follows: 39.44 for LANA, 40.52 for ORF59 and > 40 (not detectable) for K8.1. For a 40-cycle reaction, non-amplifying samples were set to Cq = 41 for the purposes of calculation. The lowest Cq value obtained in a mock infected sample was assigned as the limit of detection for each target, and data points that fall below this threshold are denoted with red shading. Yellow shading highlights values between 1.7 and 3.3 cycles lower than the limit of detection and corresponds to 5–10 fold increases in gene expression. Green shading highlights values more than 3.3 cycles lower than the limit of detection and corresponds to gene expression levels greater than 10-fold above the limit of detection. ANOVA analysis of raw Cq values revealed a statistically significant effect of KSHV infection for all target genes when comparing aggregate trends for mock vs KSHV samples over time: LANA p = 0.0006; K8.1 p = 0.02, ORF59 p

    Article Snippet: Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)).

    Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction

    A variety of B lymphocyte lineages from human tonsil are susceptible to infection with BAC16 KSHV. Magnetically sorted total B lymphocytes from four tonsil specimens were infected with KSHV or mock-infected and analyzed by FCM at indicated timepoints for (A) GFP expression and (B) immunophenotypic markers for lineage. In both cases, cells were gated for singlet/viable/CD19+. Memory B cells were further defined as CD38low/IgD-/CD27+, naïve B cells were CD38low/IgD+/CD27-, natural effector (Nat Effector) cells were CD38low/IgD+/CD27+ and germinal center (GC) cells were CD38hi/IgD-. (C) In similar infection experiments with four tonsil specimens, total RNA was extracted at 2, 4 and 6 days post-infection and viral gene transcription was verified in two technical replicates by RT-PCR. Replicate RT negative cDNA reactions for KSHV infected samples at 6 days post-infection were included as a control for DNA contamination and mean NRT Cq values (n = 8) for each target were as follows: 39.44 for LANA, 40.52 for ORF59 and > 40 (not detectable) for K8.1. For a 40-cycle reaction, non-amplifying samples were set to Cq = 41 for the purposes of calculation. The lowest Cq value obtained in a mock infected sample was assigned as the limit of detection for each target, and data points that fall below this threshold are denoted with red shading. Yellow shading highlights values between 1.7 and 3.3 cycles lower than the limit of detection and corresponds to 5–10 fold increases in gene expression. Green shading highlights values more than 3.3 cycles lower than the limit of detection and corresponds to gene expression levels greater than 10-fold above the limit of detection. ANOVA analysis of raw Cq values revealed a statistically significant effect of KSHV infection for all target genes when comparing aggregate trends for mock vs KSHV samples over time: LANA p = 0.0006; K8.1 p = 0.02, ORF59 p

    Journal: PLoS Pathogens

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes

    doi: 10.1371/journal.ppat.1006967

    Figure Lengend Snippet: A variety of B lymphocyte lineages from human tonsil are susceptible to infection with BAC16 KSHV. Magnetically sorted total B lymphocytes from four tonsil specimens were infected with KSHV or mock-infected and analyzed by FCM at indicated timepoints for (A) GFP expression and (B) immunophenotypic markers for lineage. In both cases, cells were gated for singlet/viable/CD19+. Memory B cells were further defined as CD38low/IgD-/CD27+, naïve B cells were CD38low/IgD+/CD27-, natural effector (Nat Effector) cells were CD38low/IgD+/CD27+ and germinal center (GC) cells were CD38hi/IgD-. (C) In similar infection experiments with four tonsil specimens, total RNA was extracted at 2, 4 and 6 days post-infection and viral gene transcription was verified in two technical replicates by RT-PCR. Replicate RT negative cDNA reactions for KSHV infected samples at 6 days post-infection were included as a control for DNA contamination and mean NRT Cq values (n = 8) for each target were as follows: 39.44 for LANA, 40.52 for ORF59 and > 40 (not detectable) for K8.1. For a 40-cycle reaction, non-amplifying samples were set to Cq = 41 for the purposes of calculation. The lowest Cq value obtained in a mock infected sample was assigned as the limit of detection for each target, and data points that fall below this threshold are denoted with red shading. Yellow shading highlights values between 1.7 and 3.3 cycles lower than the limit of detection and corresponds to 5–10 fold increases in gene expression. Green shading highlights values more than 3.3 cycles lower than the limit of detection and corresponds to gene expression levels greater than 10-fold above the limit of detection. ANOVA analysis of raw Cq values revealed a statistically significant effect of KSHV infection for all target genes when comparing aggregate trends for mock vs KSHV samples over time: LANA p = 0.0006; K8.1 p = 0.02, ORF59 p

    Article Snippet: Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)).

    Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction

    Biochemical characterization of RNA targeting by Cas13b. A. Thermal melting curves of PbuCas13b complexed with crRNA and target RNA. To the right of the figure legends are cartoons of the substrate used with red lines indicating mismatches (tandem mismatches are shown on the left panel; larger mismatches are shown on the right panel). Curves shifted to the left indicate a destabilized complex. B. The effect of single nucleotide mismatches between target and crRNA spacer on collateral nuclease activity activation in FLUORESCENT COLLATERAL RNA–CLEAVAGE assays. Two different guide sequences (upper, guide 1; lower, guide 2) are shown. C. . D. Diagram of the active site of PbuCas13b in the crystal structure. Citrate (shown in yellow) is bound in the active site and loop 938–951 is shown in the lower left labeled as ‘Loop’.

    Journal: Cell reports

    Article Title: High-resolution structure of Cas13b and biochemical characterization of RNA targeting and cleavage

    doi: 10.1016/j.celrep.2019.02.094

    Figure Lengend Snippet: Biochemical characterization of RNA targeting by Cas13b. A. Thermal melting curves of PbuCas13b complexed with crRNA and target RNA. To the right of the figure legends are cartoons of the substrate used with red lines indicating mismatches (tandem mismatches are shown on the left panel; larger mismatches are shown on the right panel). Curves shifted to the left indicate a destabilized complex. B. The effect of single nucleotide mismatches between target and crRNA spacer on collateral nuclease activity activation in FLUORESCENT COLLATERAL RNA–CLEAVAGE assays. Two different guide sequences (upper, guide 1; lower, guide 2) are shown. C. . D. Diagram of the active site of PbuCas13b in the crystal structure. Citrate (shown in yellow) is bound in the active site and loop 938–951 is shown in the lower left labeled as ‘Loop’.

    Article Snippet: Pre-crRNA processing was carried out in Cas13b crRNA processing buffer (10 mM TrisHCl pH 7.5, 50 mM NaCl, 0.5 mM MgCl2 , 20U SUPERase in (ThermoFisher Scientific), 0.1% BSA) for 30 minutes at 37°C, stopped by adding 2× TBE-Urea gel loading buffer and denatured for 5 minutes at 95°C.

    Techniques: Activity Assay, Activation Assay, Labeling

    PbuCas13b crRNA recognition and processing. A. Diagram of crRNA substrate co-crystallized with PbuCas13b. Direct repeat nucleotides are colored red and spacer nucleotides in light blue (full spacer is not shown). Watson-Crick base pairing denoted by black lines; non-Watson-Crick base pairing denoted by gray lines. B. The structure of crRNA within the crystallized PbuCas13b complex. The coloring is consistent with panel (A) and individual bases are numbered (−1 to −36 in the crRNA, 1 for spacer). C. Model of the 3′ end of the crRNA showing the catalytic residue K393 of the crRNA processing site and additional PbuCas13b residues that coordinate the crRNA. D. Analysis of Lid domain residues predicted to coordinate and process crRNA within PbuCas13b. Right, schematic shows Cas13b-mediated RNA degradation. The upper panel shows collateral RNase activity in FLUORESCENT COLLATERAL RNA–CLEAVAGE assays with Lid domain mutants (asterisk indicates nearly undetectable levels of fluorescence); lower panel shows processing of crRNA by these mutants. Cartoons of the expected cleavage products are shown to the left of the gel; cleavage bands and expected sizes indicated by red triangles to the right of the gel.

    Journal: Cell reports

    Article Title: High-resolution structure of Cas13b and biochemical characterization of RNA targeting and cleavage

    doi: 10.1016/j.celrep.2019.02.094

    Figure Lengend Snippet: PbuCas13b crRNA recognition and processing. A. Diagram of crRNA substrate co-crystallized with PbuCas13b. Direct repeat nucleotides are colored red and spacer nucleotides in light blue (full spacer is not shown). Watson-Crick base pairing denoted by black lines; non-Watson-Crick base pairing denoted by gray lines. B. The structure of crRNA within the crystallized PbuCas13b complex. The coloring is consistent with panel (A) and individual bases are numbered (−1 to −36 in the crRNA, 1 for spacer). C. Model of the 3′ end of the crRNA showing the catalytic residue K393 of the crRNA processing site and additional PbuCas13b residues that coordinate the crRNA. D. Analysis of Lid domain residues predicted to coordinate and process crRNA within PbuCas13b. Right, schematic shows Cas13b-mediated RNA degradation. The upper panel shows collateral RNase activity in FLUORESCENT COLLATERAL RNA–CLEAVAGE assays with Lid domain mutants (asterisk indicates nearly undetectable levels of fluorescence); lower panel shows processing of crRNA by these mutants. Cartoons of the expected cleavage products are shown to the left of the gel; cleavage bands and expected sizes indicated by red triangles to the right of the gel.

    Article Snippet: Pre-crRNA processing was carried out in Cas13b crRNA processing buffer (10 mM TrisHCl pH 7.5, 50 mM NaCl, 0.5 mM MgCl2 , 20U SUPERase in (ThermoFisher Scientific), 0.1% BSA) for 30 minutes at 37°C, stopped by adding 2× TBE-Urea gel loading buffer and denatured for 5 minutes at 95°C.

    Techniques: Activity Assay, Fluorescence

    Unique binding motifs in the N-terminus of SLBP interact with FEM1A, FEM1B, and FEM1C. (A) Diagram representing the domain structure of SLBP. The amino acid sequence and substrate receptor binding motifs representing the “degron hotspot” are shown. TAD, translational activation domain; NLS, nuclear localization sequence; RBD, RNA binding domain. (B) FEM1A, FEM1B, and FEM1C interact with amino acids 1–99 of SLBP. C-E Mapping the FEM1A, FEM1B and FEM1C binding regions in SLBP. HEK293T cells were transfected with either empty vector (EV) or FS-tagged SLBP constructs. MLN4924 was added to the cells for 4 hours before collection. Cell lysates were affinity precipitated with anti-STREP resin, and affinity precipitations were probed with the indicated antibodies. (F) The ligase-deficient SLBP(ABCdegron) mutant is unable to bind to CTIF. HEK293T cells were transfected with FLAG-tagged SLBP constructs. Cell lysates were supplemented with SUPERase-In™ RNase Inhibitor and immunoprecipitated with anti-FLAG resin. The immunoprecipitations were probed with the indicated antibodies.

    Journal: Cell Cycle

    Article Title: FEM1 proteins are ancient regulators of SLBP degradation

    doi: 10.1080/15384101.2017.1284715

    Figure Lengend Snippet: Unique binding motifs in the N-terminus of SLBP interact with FEM1A, FEM1B, and FEM1C. (A) Diagram representing the domain structure of SLBP. The amino acid sequence and substrate receptor binding motifs representing the “degron hotspot” are shown. TAD, translational activation domain; NLS, nuclear localization sequence; RBD, RNA binding domain. (B) FEM1A, FEM1B, and FEM1C interact with amino acids 1–99 of SLBP. C-E Mapping the FEM1A, FEM1B and FEM1C binding regions in SLBP. HEK293T cells were transfected with either empty vector (EV) or FS-tagged SLBP constructs. MLN4924 was added to the cells for 4 hours before collection. Cell lysates were affinity precipitated with anti-STREP resin, and affinity precipitations were probed with the indicated antibodies. (F) The ligase-deficient SLBP(ABCdegron) mutant is unable to bind to CTIF. HEK293T cells were transfected with FLAG-tagged SLBP constructs. Cell lysates were supplemented with SUPERase-In™ RNase Inhibitor and immunoprecipitated with anti-FLAG resin. The immunoprecipitations were probed with the indicated antibodies.

    Article Snippet: SUPERase-In™ RNase Inhibitor (Thermo Fisher Scientific) was used at 1U/μL where indicated.

    Techniques: Binding Assay, Sequencing, Activation Assay, RNA Binding Assay, Transfection, Plasmid Preparation, Construct, Mutagenesis, Immunoprecipitation