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Applichem rnasea
Rnasea, supplied by Applichem, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
rnasea - by Bioz Stars, 2020-09
92/100 stars

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Staining:

Article Title: Casein kinase 1α has a non-redundant and dominant role within the CK1 family in melanoma progression
Article Snippet: .. Cells were cultured for 48 h before permeabilization and fixation of the cells in 70 % ice-cold ethanol for at least 1 h. Then they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem, Darmstadt, Germany) and 50 μg/ml propidium iodide (Sigma, Taufkirchen, Germany) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phase was performed with a LSRII FACS (BD, Heidelberg, Germany) using the FACSDiva software. .. 3D Melanoma spheroid culture 2.5 × 103 SKMel19 cells were cultured on 1.5 % noble agar (Difco/BD, Heidelberg, Germany) coated 96well plates to form spheroids within 3 days.

Article Title: A Nexus Consisting of Beta-Catenin and Stat3 Attenuates BRAF Inhibitor Efficacy and Mediates Acquired Resistance to Vemurafenib
Article Snippet: .. Cells were commonly treated for 72 h. After permeabilization of the cells in 70% ice-cold ethanol for at least 1 h, they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem) and 50 μg/ml propidium iodide (Sigma) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phases was performed with a LSRII FACS (BD) using FACSDiva software (BD). .. 2.6 SA-Beta-Galactosidase Staining The Senescence Cells Histochemical Staining Kit (Sigma-Aldrich) was used according to the manufacturer's recommendations.

Incubation:

Article Title: Shikonin Directly Targets Mitochondria and Causes Mitochondrial Dysfunction in Cancer Cells
Article Snippet: .. After washing in PBS, cells were incubated with 10 μ g/mL RNaseA (Applichem Lifescience, Germany) and 50 μ g/mL propidium iodide (PI, Sigma-Aldrich, Germany) in PBS for 1 h in the dark. .. Cells were measured on an LSR-Fortessa FACS analyzer (Becton-Dickinson, Germany).

FACS:

Article Title: Casein kinase 1α has a non-redundant and dominant role within the CK1 family in melanoma progression
Article Snippet: .. Cells were cultured for 48 h before permeabilization and fixation of the cells in 70 % ice-cold ethanol for at least 1 h. Then they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem, Darmstadt, Germany) and 50 μg/ml propidium iodide (Sigma, Taufkirchen, Germany) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phase was performed with a LSRII FACS (BD, Heidelberg, Germany) using the FACSDiva software. .. 3D Melanoma spheroid culture 2.5 × 103 SKMel19 cells were cultured on 1.5 % noble agar (Difco/BD, Heidelberg, Germany) coated 96well plates to form spheroids within 3 days.

Article Title: A Nexus Consisting of Beta-Catenin and Stat3 Attenuates BRAF Inhibitor Efficacy and Mediates Acquired Resistance to Vemurafenib
Article Snippet: .. Cells were commonly treated for 72 h. After permeabilization of the cells in 70% ice-cold ethanol for at least 1 h, they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem) and 50 μg/ml propidium iodide (Sigma) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phases was performed with a LSRII FACS (BD) using FACSDiva software (BD). .. 2.6 SA-Beta-Galactosidase Staining The Senescence Cells Histochemical Staining Kit (Sigma-Aldrich) was used according to the manufacturer's recommendations.

Cell Culture:

Article Title: Casein kinase 1α has a non-redundant and dominant role within the CK1 family in melanoma progression
Article Snippet: .. Cells were cultured for 48 h before permeabilization and fixation of the cells in 70 % ice-cold ethanol for at least 1 h. Then they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem, Darmstadt, Germany) and 50 μg/ml propidium iodide (Sigma, Taufkirchen, Germany) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phase was performed with a LSRII FACS (BD, Heidelberg, Germany) using the FACSDiva software. .. 3D Melanoma spheroid culture 2.5 × 103 SKMel19 cells were cultured on 1.5 % noble agar (Difco/BD, Heidelberg, Germany) coated 96well plates to form spheroids within 3 days.

Software:

Article Title: Casein kinase 1α has a non-redundant and dominant role within the CK1 family in melanoma progression
Article Snippet: .. Cells were cultured for 48 h before permeabilization and fixation of the cells in 70 % ice-cold ethanol for at least 1 h. Then they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem, Darmstadt, Germany) and 50 μg/ml propidium iodide (Sigma, Taufkirchen, Germany) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phase was performed with a LSRII FACS (BD, Heidelberg, Germany) using the FACSDiva software. .. 3D Melanoma spheroid culture 2.5 × 103 SKMel19 cells were cultured on 1.5 % noble agar (Difco/BD, Heidelberg, Germany) coated 96well plates to form spheroids within 3 days.

Article Title: A Nexus Consisting of Beta-Catenin and Stat3 Attenuates BRAF Inhibitor Efficacy and Mediates Acquired Resistance to Vemurafenib
Article Snippet: .. Cells were commonly treated for 72 h. After permeabilization of the cells in 70% ice-cold ethanol for at least 1 h, they were re-suspended in PBS with 100 μg/ml RNAseA (Applichem) and 50 μg/ml propidium iodide (Sigma) and stained for 30 min. FACS analysis for the detection of the distribution of the cells in the each cell cycle phases was performed with a LSRII FACS (BD) using FACSDiva software (BD). .. 2.6 SA-Beta-Galactosidase Staining The Senescence Cells Histochemical Staining Kit (Sigma-Aldrich) was used according to the manufacturer's recommendations.

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    Applichem rnase a
    Dbp5 interacts with Mex67 in vivo and in vitro . (A) Dbp5 interacts mainly RNA-dependently with Mex67 in vivo . Western blot analyses of GFP-Dbp5 immunoprecipitations show strong co-precipitation of Mex67 without <t>RNase</t> A treatment, while addition of RNase A leads to a weaker, but reproducible interaction with Mex67. Detection of Hem15 served as non-binding control. (B) Direct interaction of recombinant GST-Dbp5 and Mex67, which is increased by ATP addition. Western blot analyses of Glutathione Sepharose pull-downs with GST-Dbp5 or GST and the purified heterodimer His-Mtr2 and Mex67 are shown in the presence or absence of 1 mM ATP and 0.2 mg/ml RNase A. The corresponding Coomassie-stained gel is depicted in S8D Fig .
    Rnase A, supplied by Applichem, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase a/product/Applichem
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rnase a - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

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    Dbp5 interacts with Mex67 in vivo and in vitro . (A) Dbp5 interacts mainly RNA-dependently with Mex67 in vivo . Western blot analyses of GFP-Dbp5 immunoprecipitations show strong co-precipitation of Mex67 without RNase A treatment, while addition of RNase A leads to a weaker, but reproducible interaction with Mex67. Detection of Hem15 served as non-binding control. (B) Direct interaction of recombinant GST-Dbp5 and Mex67, which is increased by ATP addition. Western blot analyses of Glutathione Sepharose pull-downs with GST-Dbp5 or GST and the purified heterodimer His-Mtr2 and Mex67 are shown in the presence or absence of 1 mM ATP and 0.2 mg/ml RNase A. The corresponding Coomassie-stained gel is depicted in S8D Fig .

    Journal: PLoS ONE

    Article Title: Nuclear Export of Pre-Ribosomal Subunits Requires Dbp5, but Not as an RNA-Helicase as for mRNA Export

    doi: 10.1371/journal.pone.0149571

    Figure Lengend Snippet: Dbp5 interacts with Mex67 in vivo and in vitro . (A) Dbp5 interacts mainly RNA-dependently with Mex67 in vivo . Western blot analyses of GFP-Dbp5 immunoprecipitations show strong co-precipitation of Mex67 without RNase A treatment, while addition of RNase A leads to a weaker, but reproducible interaction with Mex67. Detection of Hem15 served as non-binding control. (B) Direct interaction of recombinant GST-Dbp5 and Mex67, which is increased by ATP addition. Western blot analyses of Glutathione Sepharose pull-downs with GST-Dbp5 or GST and the purified heterodimer His-Mtr2 and Mex67 are shown in the presence or absence of 1 mM ATP and 0.2 mg/ml RNase A. The corresponding Coomassie-stained gel is depicted in S8D Fig .

    Article Snippet: RNase A degrades single-stranded RNAs, among them mRNAs engaged in translation, which subsequently destroys the polysomes and leads to one single 80S peak (see profiles of ).

    Techniques: In Vivo, In Vitro, Western Blot, Binding Assay, Recombinant, Purification, Staining

    Dbp5 does not displace Mex67 from exported ribosomal particles. (A) Sucrose-density fractionation experiments reveal that Mex67 is associated with polysome-containing fractions from wild type and rat8-2 cells and (B) remains equally ribosome bound upon RNase A treatment. The upper panel shows profiles of wild type and rat8-2 cells upon flow through photometry (A 254nm ) from ultra-centrifuged sucrose gradients. Before loading, the cells were shifted for 1 h to 37°C and half of the resulting lysates were treated with RNase A. The bottom panel shows the corresponding separated protein fractions in Western blot analyses with direct antibodies against Mex67 and the ribosomal protein Rps3. The mRNA-binding protein GFP-Cbp80 was expressed with a galactose-inducible promoter, subsequently detected with an anti-GFP antibody to serve as a positive control. The ratios of all proteins from the first fractions (non-ribosomal+40S+60S) and the last fractions (80S+polysomal fractions) are indicated. (C-D) Mex67 is RNA-independently associated with polysomes in wild type cells. Western blot analyses of Rpl25-GFP immunoprecipitations from polysomal gradient fractions show co-precipitation of Mex67. Wild type yeast lysates were separated in sucrose-density gradients that were subsequently fractionated. The polysomes-containing fractions (red framed area) were subjected to co-immunoprecipitation experiments that were treated with RNase A. The correct fractions were chosen according the flow through photometry profile displayed in (C) . Detection of Rps3 and Dbp5 served as positive controls. Aco1 was detected as non-binding control and the mRNA-binding protein Pab1 as control for the RNase A treatment. (E) The association of Mex67 with ribosomal proteins is unchanged in the rat8-2 strain. Western blot analyses of a co-immunoprecipitation experiment with Mex67-GFP and Rps3 or Rpl35 in wild type and rat8-2 cells, which were shifted for 1 h to 37°C, are shown upon RNase A treatment. Hem15 served as a negative control.

    Journal: PLoS ONE

    Article Title: Nuclear Export of Pre-Ribosomal Subunits Requires Dbp5, but Not as an RNA-Helicase as for mRNA Export

    doi: 10.1371/journal.pone.0149571

    Figure Lengend Snippet: Dbp5 does not displace Mex67 from exported ribosomal particles. (A) Sucrose-density fractionation experiments reveal that Mex67 is associated with polysome-containing fractions from wild type and rat8-2 cells and (B) remains equally ribosome bound upon RNase A treatment. The upper panel shows profiles of wild type and rat8-2 cells upon flow through photometry (A 254nm ) from ultra-centrifuged sucrose gradients. Before loading, the cells were shifted for 1 h to 37°C and half of the resulting lysates were treated with RNase A. The bottom panel shows the corresponding separated protein fractions in Western blot analyses with direct antibodies against Mex67 and the ribosomal protein Rps3. The mRNA-binding protein GFP-Cbp80 was expressed with a galactose-inducible promoter, subsequently detected with an anti-GFP antibody to serve as a positive control. The ratios of all proteins from the first fractions (non-ribosomal+40S+60S) and the last fractions (80S+polysomal fractions) are indicated. (C-D) Mex67 is RNA-independently associated with polysomes in wild type cells. Western blot analyses of Rpl25-GFP immunoprecipitations from polysomal gradient fractions show co-precipitation of Mex67. Wild type yeast lysates were separated in sucrose-density gradients that were subsequently fractionated. The polysomes-containing fractions (red framed area) were subjected to co-immunoprecipitation experiments that were treated with RNase A. The correct fractions were chosen according the flow through photometry profile displayed in (C) . Detection of Rps3 and Dbp5 served as positive controls. Aco1 was detected as non-binding control and the mRNA-binding protein Pab1 as control for the RNase A treatment. (E) The association of Mex67 with ribosomal proteins is unchanged in the rat8-2 strain. Western blot analyses of a co-immunoprecipitation experiment with Mex67-GFP and Rps3 or Rpl35 in wild type and rat8-2 cells, which were shifted for 1 h to 37°C, are shown upon RNase A treatment. Hem15 served as a negative control.

    Article Snippet: RNase A degrades single-stranded RNAs, among them mRNAs engaged in translation, which subsequently destroys the polysomes and leads to one single 80S peak (see profiles of ).

    Techniques: Fractionation, Flow Cytometry, Western Blot, Binding Assay, Positive Control, Immunoprecipitation, Negative Control