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Boehringer Mannheim rnasea
Rnasea, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rnasea/product/Boehringer Mannheim
Average 92 stars, based on 8 article reviews
Price from $9.99 to $1999.99
rnasea - by Bioz Stars, 2020-09
92/100 stars

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Cell Cycle Assay:

Article Title: MiR-328 Expression Is Decreased in High-Grade Gliomas and Is Associated with Worse Survival in Primary Glioblastoma
Article Snippet: .. Cell Cycle Analysis After treatment with miR-328 mimics for 48 h, cell cycle analysis was applied, in brief, cells were collected, washed with phosphate buffered saline (PBS) and then fixed with 75% ethanol at −20°C for 1.5 h, then cells were suspended in hanks balanced salt solutions (HBSS) containing 50 µg/ml of RNaseA (Boehringer Mannheim, Indianapolis, IN) and 50 µg/ml of propidium iodide (PI) (Sigma-Aldrich), incubated for 1 h at room temperature and then analyzed by FACScan (Becton Dickinson, San Jose, CA). .. Western Blotting Assay Total proteins were collected by using Total Protein Extraction Kit (KeyGen, China), 30 ug protein were added for One-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis using the discontinuous buffer system of Laemmli (Bio-Rad Laboratories, USA).

Ethanol Precipitation:

Article Title: The complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans
Article Snippet: .. The DNA was treated with RNaseA (Boehringer Mannheim Biochemicals) and purified by phenol–chloroform extraction and ethanol precipitation. ..

Purification:

Article Title: The complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans
Article Snippet: .. The DNA was treated with RNaseA (Boehringer Mannheim Biochemicals) and purified by phenol–chloroform extraction and ethanol precipitation. ..

Incubation:

Article Title: Telomere loss in cells treated with cisplatin
Article Snippet: .. The cells were then incubated in 1 ml PBS containing 50 μg/ml propidium iodide (Sigma) and 250 μg/ml RNaseA (Boehringer Mannheim) at 37°C for 30 min to stain the DNA and eliminate RNA. .. The fluorescence intensity in the cell was measured by a FACScan (Becton Dickinson).

Article Title: Synchronous Onset of NGF and TrkA Survival Dependence in Developing Dorsal Root Ganglia
Article Snippet: .. Sections were incubated with individual sense-strand33 P-labeled riboprobes or were pretreated with RNaseA (Boehringer Mannheim, Mannheim, Germany) (20 μg/ml for 30 min at 37°C), followed by hybridization with individual antisense riboprobes. .. In each case, control hybridizations resulted in loss of the specific hybridization patterns noted with the complementary RNA probes.

Article Title: MiR-328 Expression Is Decreased in High-Grade Gliomas and Is Associated with Worse Survival in Primary Glioblastoma
Article Snippet: .. Cell Cycle Analysis After treatment with miR-328 mimics for 48 h, cell cycle analysis was applied, in brief, cells were collected, washed with phosphate buffered saline (PBS) and then fixed with 75% ethanol at −20°C for 1.5 h, then cells were suspended in hanks balanced salt solutions (HBSS) containing 50 µg/ml of RNaseA (Boehringer Mannheim, Indianapolis, IN) and 50 µg/ml of propidium iodide (PI) (Sigma-Aldrich), incubated for 1 h at room temperature and then analyzed by FACScan (Becton Dickinson, San Jose, CA). .. Western Blotting Assay Total proteins were collected by using Total Protein Extraction Kit (KeyGen, China), 30 ug protein were added for One-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis using the discontinuous buffer system of Laemmli (Bio-Rad Laboratories, USA).

Article Title: Graft-versus-host-disease-associated donor cell engraftment in an F1 hybrid model is dependent upon the Fas pathway
Article Snippet: .. The samples were incubated with 200 µg/ml of RNaseA (Boehringer Mannheim, Indianapolis, IN) for 1 hr at 37° and then with 400 µg/ml of proteinase K (Wako Pure Chemical Industries) for 2 hr at 60°. ..

Modification:

Article Title: Plasmid copy-number control and better-than-random segregation genes of pSM19035 share a common regulator
Article Snippet: .. DNA restriction and modification enzymes, ATP, RNaseA, and poly[d(I/C)] were purchased from Boehringer Mannheim. .. Gel-purified DNA fragments were end labeled as described ( ).

Staining:

Article Title: Telomere loss in cells treated with cisplatin
Article Snippet: .. The cells were then incubated in 1 ml PBS containing 50 μg/ml propidium iodide (Sigma) and 250 μg/ml RNaseA (Boehringer Mannheim) at 37°C for 30 min to stain the DNA and eliminate RNA. .. The fluorescence intensity in the cell was measured by a FACScan (Becton Dickinson).

Hybridization:

Article Title: Synchronous Onset of NGF and TrkA Survival Dependence in Developing Dorsal Root Ganglia
Article Snippet: .. Sections were incubated with individual sense-strand33 P-labeled riboprobes or were pretreated with RNaseA (Boehringer Mannheim, Mannheim, Germany) (20 μg/ml for 30 min at 37°C), followed by hybridization with individual antisense riboprobes. .. In each case, control hybridizations resulted in loss of the specific hybridization patterns noted with the complementary RNA probes.

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    Boehringer Mannheim l rnase t1
    Activity of a bimolecular twister sister ribozyme (a) Sequence and secondary structure model of a bimolecular construct derived from TS-1 ( Supplementary Fig. 1 ). Highly conserved nucleotides are depicted in red and Clv designates the cleavage site. Lowercase letters identify non-native guanosine residues that were added to facilitate in vitro transcription. (b) PAGE separation of bimolecular TS-1 ribozyme assay products demonstrating the requirement for the enzyme strand and for divalent metal ions. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate and 5′ Clv identifies the cleavage product. Reactions were conducted as described (see Online Methods ) with variations noted. (c) PAGE separation of ribozyme cleavage products. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate, and C13 identifies the 5′ 32 P-labeled fragment band produced by incubation with excess unlabeled ribozyme for 30 min (Rxn lane). NR, T1 and − OH lanes designate no reaction, <t>RNase</t> T1 partial digestion (cleaves after G nucleotides), or partial alkaline digestion (cleaves all internucleotides), respectively. (d) Mass spectrum analysis of bimolecular TS-1 cleavage reaction products were examined by mass spectrometry (see Online Methods ). The second largest peak near the 5′ Clv annotation is the spontaneously formed opened version (observed mass, 4304.576) of the initial 2′,3′-cyclic phosphate product. Intensity is abbreviated int. (e) The dependence of twister sister rate constants on pH. (f) The dependence of Mg 2+ concentration on twister sister rate constants. A version of this figure containing full-length gel images is shown in Supplementary Figure 11 .
    L Rnase T1, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/l rnase t1/product/Boehringer Mannheim
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    92
    Boehringer Mannheim rnase h
    Polyadenylylation in polysomes occurs on  mRNA and not rRNA. RNA was isolated and treated as described. Following  electrophoresis on 6% denaturing PAGE, the Northern blots were probed  with a full-length  lpp  probe ( A ) or an  oligonucleotide complementary to the 3′ end of the mature 23S rRNA  ( B ). Exogenous PAP was added as indicated at 0.4  units/reaction. All samples were digested with RNase H.
    Rnase H, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase h/product/Boehringer Mannheim
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    85
    Boehringer Mannheim monoclonal antibody against rnase l
    E3L of VV evades the IFN system by blocking IFN induction through IRF3 and IFN action through the 2-5A/RNase L pathway and protein kinase PKR.
    Monoclonal Antibody Against Rnase L, supplied by Boehringer Mannheim, 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/monoclonal antibody against rnase l/product/Boehringer Mannheim
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    93
    Boehringer Mannheim rnase a
    Qk1 self-associates into homodimers in the absence of cellular RNA. (A) C6 glioma cells were treated in situ with (+) or without (−) BMH. The cells were lysed in sample buffer, and the proteins were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with rabbit anti-Qk1 antibody. The bands representing the three Qk1 isoforms are shown as monomers, and the cross-linked complexes are indicated. The positions of molecular mass markers (in kilodaltons) are indicated on the left. (B) HeLa cells cotransfected with myc- and HA-Qk1 were treated in situ with BMH. The cells were lysed, and an aliquot of the total cell lysate (TCL) as well as anti-myc (myc) and IgG (C) immunoprecipitates were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with anti-HA antibodies. (C) Qk1, unlike GRP33, does not require cellular RNA for self-association. HeLa cells expressing myc-Qk1, HA-Qk1, myc-GRP33, or HA-GRP33 were lysed separately, and each cell lysate was divided into two portions, either treated with <t>RNase</t> A (+) or not treated (−), mixed, and incubated with anti-myc antibodies. The anti-myc immunoprecipitates (IP) of the mixed lysates were separated by SDS-PAGE and immunoblotted with anti-HA antibodies.
    Rnase A, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 93/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Activity of a bimolecular twister sister ribozyme (a) Sequence and secondary structure model of a bimolecular construct derived from TS-1 ( Supplementary Fig. 1 ). Highly conserved nucleotides are depicted in red and Clv designates the cleavage site. Lowercase letters identify non-native guanosine residues that were added to facilitate in vitro transcription. (b) PAGE separation of bimolecular TS-1 ribozyme assay products demonstrating the requirement for the enzyme strand and for divalent metal ions. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate and 5′ Clv identifies the cleavage product. Reactions were conducted as described (see Online Methods ) with variations noted. (c) PAGE separation of ribozyme cleavage products. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate, and C13 identifies the 5′ 32 P-labeled fragment band produced by incubation with excess unlabeled ribozyme for 30 min (Rxn lane). NR, T1 and − OH lanes designate no reaction, RNase T1 partial digestion (cleaves after G nucleotides), or partial alkaline digestion (cleaves all internucleotides), respectively. (d) Mass spectrum analysis of bimolecular TS-1 cleavage reaction products were examined by mass spectrometry (see Online Methods ). The second largest peak near the 5′ Clv annotation is the spontaneously formed opened version (observed mass, 4304.576) of the initial 2′,3′-cyclic phosphate product. Intensity is abbreviated int. (e) The dependence of twister sister rate constants on pH. (f) The dependence of Mg 2+ concentration on twister sister rate constants. A version of this figure containing full-length gel images is shown in Supplementary Figure 11 .

    Journal: Nature chemical biology

    Article Title: New classes of self-cleaving ribozymes revealed by comparative genomics analysis

    doi: 10.1038/nchembio.1846

    Figure Lengend Snippet: Activity of a bimolecular twister sister ribozyme (a) Sequence and secondary structure model of a bimolecular construct derived from TS-1 ( Supplementary Fig. 1 ). Highly conserved nucleotides are depicted in red and Clv designates the cleavage site. Lowercase letters identify non-native guanosine residues that were added to facilitate in vitro transcription. (b) PAGE separation of bimolecular TS-1 ribozyme assay products demonstrating the requirement for the enzyme strand and for divalent metal ions. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate and 5′ Clv identifies the cleavage product. Reactions were conducted as described (see Online Methods ) with variations noted. (c) PAGE separation of ribozyme cleavage products. S designates the 5′ 32 P-labeled 23-nucleotide RNA substrate, and C13 identifies the 5′ 32 P-labeled fragment band produced by incubation with excess unlabeled ribozyme for 30 min (Rxn lane). NR, T1 and − OH lanes designate no reaction, RNase T1 partial digestion (cleaves after G nucleotides), or partial alkaline digestion (cleaves all internucleotides), respectively. (d) Mass spectrum analysis of bimolecular TS-1 cleavage reaction products were examined by mass spectrometry (see Online Methods ). The second largest peak near the 5′ Clv annotation is the spontaneously formed opened version (observed mass, 4304.576) of the initial 2′,3′-cyclic phosphate product. Intensity is abbreviated int. (e) The dependence of twister sister rate constants on pH. (f) The dependence of Mg 2+ concentration on twister sister rate constants. A version of this figure containing full-length gel images is shown in Supplementary Figure 11 .

    Article Snippet: To prepare the RNA marker lanes, the radiolabeled substrate was partially digested with RNase T1 nuclease [25 mM sodium citrate (pH 5.0 at 23°C), 4 M urea, 0.6 mM EDTA, and 0.2 U per L RNase T1 (from Aspergillus oryzae ; Boehringer Mannheim) for 11 min at 55°C] or with alkali [50 mM Na2 CO3 (pH 9.0 at 23°C) and 1 mM EDTA for 7 min at 90°C].

    Techniques: Activity Assay, Sequencing, Construct, Derivative Assay, In Vitro, Polyacrylamide Gel Electrophoresis, Labeling, Produced, Incubation, Mass Spectrometry, Concentration Assay

    Polyadenylylation in polysomes occurs on  mRNA and not rRNA. RNA was isolated and treated as described. Following  electrophoresis on 6% denaturing PAGE, the Northern blots were probed  with a full-length  lpp  probe ( A ) or an  oligonucleotide complementary to the 3′ end of the mature 23S rRNA  ( B ). Exogenous PAP was added as indicated at 0.4  units/reaction. All samples were digested with RNase H.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Development of an in vitro mRNA decay system for Escherichia coli: Poly(A) polymerase I is necessary to trigger degradation

    doi:

    Figure Lengend Snippet: Polyadenylylation in polysomes occurs on mRNA and not rRNA. RNA was isolated and treated as described. Following electrophoresis on 6% denaturing PAGE, the Northern blots were probed with a full-length lpp probe ( A ) or an oligonucleotide complementary to the 3′ end of the mature 23S rRNA ( B ). Exogenous PAP was added as indicated at 0.4 units/reaction. All samples were digested with RNase H.

    Article Snippet: Nucleotides, RNase H, RNase T1, RNase A, and T4 RNA ligase were purchased from Boehringer Mannheim.

    Techniques: Isolation, Electrophoresis, Polyacrylamide Gel Electrophoresis, Northern Blot

    E3L of VV evades the IFN system by blocking IFN induction through IRF3 and IFN action through the 2-5A/RNase L pathway and protein kinase PKR.

    Journal: Journal of Virology

    Article Title: Blockade of Interferon Induction and Action by the E3L Double-Stranded RNA Binding Proteins of Vaccinia Virus

    doi: 10.1128/JVI.76.10.5251-5259.2002

    Figure Lengend Snippet: E3L of VV evades the IFN system by blocking IFN induction through IRF3 and IFN action through the 2-5A/RNase L pathway and protein kinase PKR.

    Article Snippet: Proteins (200 μg per lane) in cell extracts were separated on 10% polyacrylamide/sodium dodecyl sulfate (SDS) gels, transferred to Immobilon-P membrane (Millipore), and incubated with monoclonal antibody against RNase L ( ) or monoclonal antibody against β-actin (Boehringer Mannheim) for 1 h. Membranes were washed with PBS containing 0.1% Tween 20 and incubated with goat anti-mouse antibody tagged with horseradish peroxidase (Gibco BRL) for 1 h. Proteins were visualized by enhanced chemiluminescence (ECL) reagent (Amersham).

    Techniques: Blocking Assay

    Impact of RNase L and PKR deficiencies on viral growth and pathogenesis. (A) One-step growth of VV and VVΔE3L (Copenhagen strain) in wild-type and RNase L −/− PKR −/− cells. Cells were infected at an MOI of 6. At different times postinfection, viruses were harvested and virus yields were determined by plaque titration on BHK21 cells. (B) VV but not VVΔE3L is lethal for mice regardless of the presence or absence of RNase L and PKR. Seven-week-old wild-type, RNase L −/− , PKR −/− , and RNase L −/− PKR −/− mice were injected i.n. with 10 6 PFU of wild-type VV or 5 × 10 6 PFU of VVΔE3L (WR strain). Survival was monitored daily, and the mouse survival was plotted.

    Journal: Journal of Virology

    Article Title: Blockade of Interferon Induction and Action by the E3L Double-Stranded RNA Binding Proteins of Vaccinia Virus

    doi: 10.1128/JVI.76.10.5251-5259.2002

    Figure Lengend Snippet: Impact of RNase L and PKR deficiencies on viral growth and pathogenesis. (A) One-step growth of VV and VVΔE3L (Copenhagen strain) in wild-type and RNase L −/− PKR −/− cells. Cells were infected at an MOI of 6. At different times postinfection, viruses were harvested and virus yields were determined by plaque titration on BHK21 cells. (B) VV but not VVΔE3L is lethal for mice regardless of the presence or absence of RNase L and PKR. Seven-week-old wild-type, RNase L −/− , PKR −/− , and RNase L −/− PKR −/− mice were injected i.n. with 10 6 PFU of wild-type VV or 5 × 10 6 PFU of VVΔE3L (WR strain). Survival was monitored daily, and the mouse survival was plotted.

    Article Snippet: Proteins (200 μg per lane) in cell extracts were separated on 10% polyacrylamide/sodium dodecyl sulfate (SDS) gels, transferred to Immobilon-P membrane (Millipore), and incubated with monoclonal antibody against RNase L ( ) or monoclonal antibody against β-actin (Boehringer Mannheim) for 1 h. Membranes were washed with PBS containing 0.1% Tween 20 and incubated with goat anti-mouse antibody tagged with horseradish peroxidase (Gibco BRL) for 1 h. Proteins were visualized by enhanced chemiluminescence (ECL) reagent (Amersham).

    Techniques: Infection, Titration, Mouse Assay, Injection

    E3L allows VV to evade the antiviral activity of RNase L. Plaque assays (A) and one-step viral-growth curves (B) of wild-type VV and VVΔE3L (Copenhagen strain) are shown. (C) Diagram of RNase L and mutant RNase L ΔEN . (D) Ponasterone induction of RNase L in comparison to β-actin as determined by probing a Western blot with antibodies. (E) Assay for RNase L and mutant RNase L ΔEN levels by covalent cross-linking to a 32 P-labeled 2-5A analog before and after ponasterone induction (5 μM for 24 h). (F) Assay for RNase L activity in intact cells as measured by specific cleavages in rRNAs. Cells were incubated in the absence or presence of ponasterone (5 μM) for 24 h and transfected with 2-5A [p 3 A(2′p5′A) 3 ] (1 μM) for 3 h. Total RNA was isolated, electrophoresed in a formaldehyde-1.2% agarose gel, stained with ethidium bromide (lower panel), and probed with 32 P-labeled 18S rRNA cDNA (upper panel). (G) RNase L −/− cells containing inducible cDNAs for RNase L and RNase L ΔEN were incubated in the absence or presence of ponasterone (5 μM) for 24 h and infected at an MOI of 6 with wild-type VV or VVΔE3L (Copenhagen strain). At 24 h postinfection, viruses were harvested and virus yields were determined by plaque titration on BHK21 cells.

    Journal: Journal of Virology

    Article Title: Blockade of Interferon Induction and Action by the E3L Double-Stranded RNA Binding Proteins of Vaccinia Virus

    doi: 10.1128/JVI.76.10.5251-5259.2002

    Figure Lengend Snippet: E3L allows VV to evade the antiviral activity of RNase L. Plaque assays (A) and one-step viral-growth curves (B) of wild-type VV and VVΔE3L (Copenhagen strain) are shown. (C) Diagram of RNase L and mutant RNase L ΔEN . (D) Ponasterone induction of RNase L in comparison to β-actin as determined by probing a Western blot with antibodies. (E) Assay for RNase L and mutant RNase L ΔEN levels by covalent cross-linking to a 32 P-labeled 2-5A analog before and after ponasterone induction (5 μM for 24 h). (F) Assay for RNase L activity in intact cells as measured by specific cleavages in rRNAs. Cells were incubated in the absence or presence of ponasterone (5 μM) for 24 h and transfected with 2-5A [p 3 A(2′p5′A) 3 ] (1 μM) for 3 h. Total RNA was isolated, electrophoresed in a formaldehyde-1.2% agarose gel, stained with ethidium bromide (lower panel), and probed with 32 P-labeled 18S rRNA cDNA (upper panel). (G) RNase L −/− cells containing inducible cDNAs for RNase L and RNase L ΔEN were incubated in the absence or presence of ponasterone (5 μM) for 24 h and infected at an MOI of 6 with wild-type VV or VVΔE3L (Copenhagen strain). At 24 h postinfection, viruses were harvested and virus yields were determined by plaque titration on BHK21 cells.

    Article Snippet: Proteins (200 μg per lane) in cell extracts were separated on 10% polyacrylamide/sodium dodecyl sulfate (SDS) gels, transferred to Immobilon-P membrane (Millipore), and incubated with monoclonal antibody against RNase L ( ) or monoclonal antibody against β-actin (Boehringer Mannheim) for 1 h. Membranes were washed with PBS containing 0.1% Tween 20 and incubated with goat anti-mouse antibody tagged with horseradish peroxidase (Gibco BRL) for 1 h. Proteins were visualized by enhanced chemiluminescence (ECL) reagent (Amersham).

    Techniques: Activity Assay, Mutagenesis, Western Blot, Labeling, Incubation, Transfection, Isolation, Agarose Gel Electrophoresis, Staining, Infection, Titration

    Qk1 self-associates into homodimers in the absence of cellular RNA. (A) C6 glioma cells were treated in situ with (+) or without (−) BMH. The cells were lysed in sample buffer, and the proteins were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with rabbit anti-Qk1 antibody. The bands representing the three Qk1 isoforms are shown as monomers, and the cross-linked complexes are indicated. The positions of molecular mass markers (in kilodaltons) are indicated on the left. (B) HeLa cells cotransfected with myc- and HA-Qk1 were treated in situ with BMH. The cells were lysed, and an aliquot of the total cell lysate (TCL) as well as anti-myc (myc) and IgG (C) immunoprecipitates were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with anti-HA antibodies. (C) Qk1, unlike GRP33, does not require cellular RNA for self-association. HeLa cells expressing myc-Qk1, HA-Qk1, myc-GRP33, or HA-GRP33 were lysed separately, and each cell lysate was divided into two portions, either treated with RNase A (+) or not treated (−), mixed, and incubated with anti-myc antibodies. The anti-myc immunoprecipitates (IP) of the mixed lysates were separated by SDS-PAGE and immunoblotted with anti-HA antibodies.

    Journal: Molecular and Cellular Biology

    Article Title: Structure-Function Analysis of Qk1: a Lethal Point Mutation in Mouse quaking Prevents Homodimerization

    doi:

    Figure Lengend Snippet: Qk1 self-associates into homodimers in the absence of cellular RNA. (A) C6 glioma cells were treated in situ with (+) or without (−) BMH. The cells were lysed in sample buffer, and the proteins were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with rabbit anti-Qk1 antibody. The bands representing the three Qk1 isoforms are shown as monomers, and the cross-linked complexes are indicated. The positions of molecular mass markers (in kilodaltons) are indicated on the left. (B) HeLa cells cotransfected with myc- and HA-Qk1 were treated in situ with BMH. The cells were lysed, and an aliquot of the total cell lysate (TCL) as well as anti-myc (myc) and IgG (C) immunoprecipitates were separated by SDS-PAGE. The proteins were transferred to nitrocellulose and immunoblotted with anti-HA antibodies. (C) Qk1, unlike GRP33, does not require cellular RNA for self-association. HeLa cells expressing myc-Qk1, HA-Qk1, myc-GRP33, or HA-GRP33 were lysed separately, and each cell lysate was divided into two portions, either treated with RNase A (+) or not treated (−), mixed, and incubated with anti-myc antibodies. The anti-myc immunoprecipitates (IP) of the mixed lysates were separated by SDS-PAGE and immunoblotted with anti-HA antibodies.

    Article Snippet: Incubating the cell lysates at 37°C without RNase A was considered a mock treatment.

    Techniques: In Situ, SDS Page, Expressing, Incubation