lysis buffer  (Millipore)


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    Name:
    2X Cell Lysis Buffer
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    Catalog Number:
    rablysis1
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    Structured Review

    Millipore lysis buffer

    https://www.bioz.com/result/lysis buffer/product/Millipore
    Average 99 stars, based on 15588 article reviews
    Price from $9.99 to $1999.99
    lysis buffer - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Transfection:

    Article Title: Histone Deacetylase 7 Associates With Runx2 and Represses Its Activity During Osteoblast Maturation in a Deacetylation-Independent Manner
    Article Snippet: .. Twenty-four hours after transfection, whole cell lysates were prepared in lysis buffer as above and immunoprecipitated with FLAG-M2-Agarose (Sigma). .. ROS17/2.8 cell nuclear lysates were prepared as for immunoprecipitation.

    Protease Inhibitor:

    Article Title: Effect of Vitamin D Supplementation on Mycobacterium tuberculosis-Induced Innate Immune Responses in a Canadian Den? First Nations Cohort
    Article Snippet: .. Western Blots Total cell lysates were prepared in lysis buffer containing 10 mM Tris-HCl pH 7.5, 150 mM NaCl, 2 mM EDTA, protease inhibitor cocktail (Sigma-Aldrich) and 1% Triton X-100. .. The lysates were electrophoretically resolved on 4–12% NuPAGE® Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Millipore, Canada).

    Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
    Article Snippet: .. Cell lysates were prepared in lysis buffer consisting of 10% glycerol, 25 m m Tris HCl, pH 7.5, 150 m m NaCl, 1% Triton X100, 5 m m EDTA, and 1 m m EGTA supplemented with 1:100 Protease Inhibitor Mixture (Sigma-Aldrich, St. Louis, MO). .. Samples were homogenized, sonicated, and centrifuged (15 min at 4°C max speed Biofuge).

    Article Title: The long noncoding RNA, treRNA, decreases DNA damage and is associated with poor response to chemotherapy in chronic lymphocytic leukemia
    Article Snippet: .. Immunoblot Whole cell lysates were prepared by lysing PBS-washed OSU-CLL cell pellets in cold lysis buffer containing phosphatase inhibitor cocktail 1 and 2, protease inhibitor cocktail P8340 and 1mM phenylmethylsulfonyl fluoride (all from Sigma). .. Protein was quantified by the BCA method (Pierce).

    Article Title: PCB153-Induced Overexpression of ID3 Contributes to the Development of Microvascular Lesions
    Article Snippet: .. Immunoblotting Whole cell lysates were prepared with lysis buffer containing [25 mM Tris-HCl buffer (pH 8.0), 150 mM NaCl, 0.2% NP-40, 10% glycerol, 8 mM β-glycerophosphate, 2.5 mM sodium pyrophosphate, 10 mM NaF, 0.2 mM Na3 VO4 , 1 mM DTT and 10 µl/ml protease inhibitor cocktail (Sigma Aldrich). .. Proteins were quantified using the Bradford Assay Reagent (Bio-Rad) according to the manufacturer's instructions.

    Immunoprecipitation:

    Article Title: Histone Deacetylase 7 Associates With Runx2 and Represses Its Activity During Osteoblast Maturation in a Deacetylation-Independent Manner
    Article Snippet: .. Twenty-four hours after transfection, whole cell lysates were prepared in lysis buffer as above and immunoprecipitated with FLAG-M2-Agarose (Sigma). .. ROS17/2.8 cell nuclear lysates were prepared as for immunoprecipitation.

    Incubation:

    Article Title: Activation of FADD-Dependent Neuronal Death Pathways as a Predictor of Pathogenicity for LRRK2 Mutations
    Article Snippet: .. In some experiments, 500 μg of clarified lysate was incubated overnight with Flag-conjugated resin (Biotool; Munich, DE), followed by washing in lysis buffer, before final elution of the protein in 2X SDS sample buffer at 95°C for 5 min. Eluate was separated by 10% SDS-PAGE, transferred to nitrocellulose, and the membranes probed with antibodies to V5 or Flag (Sigma). .. In parallel experiments, 15 μg of clarified lysate was bound to Flag-coated ELISA plates (as in the kinase activity experiments) for 2h at 37°C.

    Polyacrylamide Gel Electrophoresis:

    Article Title: LncRNA MIR100HG promotes cell proliferation in triple-negative breast cancer through triplex formation with p27 loci
    Article Snippet: .. Cell were treated with lysis buffer for 30 min. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenefluoride (PVDF) membranes (Millipore). .. The membranes were probed with specific antibodies against GAPDH (Abcam), cyclin D1 (Abcam), p21 (Abcam), and p27 (Abcam), then incubated for 1 h at room temperature with appropriate HRP-linked secondary antibodies (Sigma) and detected with chemiluminescent HRP substrate (Millipore).

    Western Blot:

    Article Title: Effect of Vitamin D Supplementation on Mycobacterium tuberculosis-Induced Innate Immune Responses in a Canadian Den? First Nations Cohort
    Article Snippet: .. Western Blots Total cell lysates were prepared in lysis buffer containing 10 mM Tris-HCl pH 7.5, 150 mM NaCl, 2 mM EDTA, protease inhibitor cocktail (Sigma-Aldrich) and 1% Triton X-100. .. The lysates were electrophoretically resolved on 4–12% NuPAGE® Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Millipore, Canada).

    Article Title: Role of the P2X7 receptor in in vitro and in vivo glioma tumor growth
    Article Snippet: .. Western blot Total U138-wt, U138-P2X7 polyclone and U138-P2X7 clone cell lysates were prepared in lysis buffer (300 μM sucrose, 1.0 mM K2 HPO4 , 5.5 mM glucose, 20 mM HEPES) supplemented with protease inhibitors (1 mM phenylmethylsulfonyl fluoride, 1 mM benzamidine, all from Sigma-Aldrich). ..

    Lysis:

    Article Title: Effect of Vitamin D Supplementation on Mycobacterium tuberculosis-Induced Innate Immune Responses in a Canadian Den? First Nations Cohort
    Article Snippet: .. Western Blots Total cell lysates were prepared in lysis buffer containing 10 mM Tris-HCl pH 7.5, 150 mM NaCl, 2 mM EDTA, protease inhibitor cocktail (Sigma-Aldrich) and 1% Triton X-100. .. The lysates were electrophoretically resolved on 4–12% NuPAGE® Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Millipore, Canada).

    Article Title: Role of the P2X7 receptor in in vitro and in vivo glioma tumor growth
    Article Snippet: .. Western blot Total U138-wt, U138-P2X7 polyclone and U138-P2X7 clone cell lysates were prepared in lysis buffer (300 μM sucrose, 1.0 mM K2 HPO4 , 5.5 mM glucose, 20 mM HEPES) supplemented with protease inhibitors (1 mM phenylmethylsulfonyl fluoride, 1 mM benzamidine, all from Sigma-Aldrich). ..

    Article Title: Widespread Disruption of Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor Occupancy at Its Target Genes in Huntington's Disease
    Article Snippet: .. Cell lysates were prepared in lysis buffer consisting of 10% glycerol, 25 m m Tris HCl, pH 7.5, 150 m m NaCl, 1% Triton X100, 5 m m EDTA, and 1 m m EGTA supplemented with 1:100 Protease Inhibitor Mixture (Sigma-Aldrich, St. Louis, MO). .. Samples were homogenized, sonicated, and centrifuged (15 min at 4°C max speed Biofuge).

    Article Title: Histone Deacetylase 7 Associates With Runx2 and Represses Its Activity During Osteoblast Maturation in a Deacetylation-Independent Manner
    Article Snippet: .. Twenty-four hours after transfection, whole cell lysates were prepared in lysis buffer as above and immunoprecipitated with FLAG-M2-Agarose (Sigma). .. ROS17/2.8 cell nuclear lysates were prepared as for immunoprecipitation.

    Article Title: The long noncoding RNA, treRNA, decreases DNA damage and is associated with poor response to chemotherapy in chronic lymphocytic leukemia
    Article Snippet: .. Immunoblot Whole cell lysates were prepared by lysing PBS-washed OSU-CLL cell pellets in cold lysis buffer containing phosphatase inhibitor cocktail 1 and 2, protease inhibitor cocktail P8340 and 1mM phenylmethylsulfonyl fluoride (all from Sigma). .. Protein was quantified by the BCA method (Pierce).

    Article Title: LncRNA MIR100HG promotes cell proliferation in triple-negative breast cancer through triplex formation with p27 loci
    Article Snippet: .. Cell were treated with lysis buffer for 30 min. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenefluoride (PVDF) membranes (Millipore). .. The membranes were probed with specific antibodies against GAPDH (Abcam), cyclin D1 (Abcam), p21 (Abcam), and p27 (Abcam), then incubated for 1 h at room temperature with appropriate HRP-linked secondary antibodies (Sigma) and detected with chemiluminescent HRP substrate (Millipore).

    Article Title: PCB153-Induced Overexpression of ID3 Contributes to the Development of Microvascular Lesions
    Article Snippet: .. Immunoblotting Whole cell lysates were prepared with lysis buffer containing [25 mM Tris-HCl buffer (pH 8.0), 150 mM NaCl, 0.2% NP-40, 10% glycerol, 8 mM β-glycerophosphate, 2.5 mM sodium pyrophosphate, 10 mM NaF, 0.2 mM Na3 VO4 , 1 mM DTT and 10 µl/ml protease inhibitor cocktail (Sigma Aldrich). .. Proteins were quantified using the Bradford Assay Reagent (Bio-Rad) according to the manufacturer's instructions.

    Article Title: Activation of FADD-Dependent Neuronal Death Pathways as a Predictor of Pathogenicity for LRRK2 Mutations
    Article Snippet: .. In some experiments, 500 μg of clarified lysate was incubated overnight with Flag-conjugated resin (Biotool; Munich, DE), followed by washing in lysis buffer, before final elution of the protein in 2X SDS sample buffer at 95°C for 5 min. Eluate was separated by 10% SDS-PAGE, transferred to nitrocellulose, and the membranes probed with antibodies to V5 or Flag (Sigma). .. In parallel experiments, 15 μg of clarified lysate was bound to Flag-coated ELISA plates (as in the kinase activity experiments) for 2h at 37°C.

    SDS Page:

    Article Title: LncRNA MIR100HG promotes cell proliferation in triple-negative breast cancer through triplex formation with p27 loci
    Article Snippet: .. Cell were treated with lysis buffer for 30 min. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenefluoride (PVDF) membranes (Millipore). .. The membranes were probed with specific antibodies against GAPDH (Abcam), cyclin D1 (Abcam), p21 (Abcam), and p27 (Abcam), then incubated for 1 h at room temperature with appropriate HRP-linked secondary antibodies (Sigma) and detected with chemiluminescent HRP substrate (Millipore).

    Article Title: Activation of FADD-Dependent Neuronal Death Pathways as a Predictor of Pathogenicity for LRRK2 Mutations
    Article Snippet: .. In some experiments, 500 μg of clarified lysate was incubated overnight with Flag-conjugated resin (Biotool; Munich, DE), followed by washing in lysis buffer, before final elution of the protein in 2X SDS sample buffer at 95°C for 5 min. Eluate was separated by 10% SDS-PAGE, transferred to nitrocellulose, and the membranes probed with antibodies to V5 or Flag (Sigma). .. In parallel experiments, 15 μg of clarified lysate was bound to Flag-coated ELISA plates (as in the kinase activity experiments) for 2h at 37°C.

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  • 99
    Millipore rna lysis buffer
    CircCLK3 expression is relatively highly expressed in cervical cancer tissues and predicts poor prognosis. a CircRNA sequencing was adapted to detect the differentially expressed circRNAs between cervical cancer tissues and matched normal tissues. b The genomic loci of the CLK3 gene and circCLK3. Sanger sequencing confirmed the head-to-hail splicing. c The expression level of circCLK3 in 5 cervical cancer cell lines relative to HeLa cell. d The gel electrophoresis validated the existence of circCLK3. Divergent primers amplified circCLK3 in cDNA but not gDNA. e qRT-PCR results of U6, GAPDH, and circCLK3 expressions in cell nuclei and cytoplasm in <t>SiHa</t> and HeLa cells. f <t>RNA-FISH</t> assays were performed to identify the subcellular location of circCLK3 with the cy3-labeled circCLK3 probe. The results showed that most of circCLK3 was located in cytoplasm in SiHa and HeLa cells. g qRT-PCR results of circCLK3 and CLK3 expression after treatment with actinomycin D at the indicated time points in SiHa and HeLa cells. h qRT-PCR results of circCLK3 and CLK3 expression after treatment with RNase R in SiHa and HeLa cells. i CircCLK3 expression was significantly higher in 48 paired fresh frozen cervical cancer tissues compared with adjacent normal tissues. j , k Kaplan–Meier Plotter analysis of the correlation between circCLK3 level with overall survive or disease-free survive of cervical cancer patients. Data are reported as means ± standard deviation of three independent experiments. * p
    Rna Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna lysis buffer/product/Millipore
    Average 99 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    rna lysis buffer - by Bioz Stars, 2020-09
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    86
    Millipore co immunoprecipitation lysis buffer
    PIWIL1 cooperates with UPF1 to negatively regulate PIWIL1-bound RNAs in a piRNA-independent manner. (A) A Coomassie blue staining protein gel of <t>PIWIL1-co-immunoprecipitation,</t> Mass spectrometry of this protein gel identifies PIWIL1 (blue arrow) as well as UPF1 (red arrow). (B) Western blotting showing reciprocal co-immunoprecipitated between PIWIL1 and UPF1 (total and phosphorylated form, p-UPF1) and between PIWIL1 and UPF2. (C) Western blotting shows that PIWIL1 co-immunoprecipitates with NMD complex core proteins UPF1, phosphorylated UPF1 (p-UPF1), UPF2, and SMG1. (D) Immunofluorescence staining of PIWIL1 (green) and DCP1A (red, a P body marker) in wildtype and PIWIL_KO SNU-1 cells. (E) Immunofluorescence staining of PIWIL1 (green), DCP1A (red), and UPF1 (fuchsia) in SNU-1 cells, which shows PIWIL1 co-localized with UPF1 in the P body. (F) Venn diagram of PIWIL1-bound RNAs, UPF1-bound RNAs, and PIWIL1-negatively regulated RNAs, with P
    Co Immunoprecipitation Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/co immunoprecipitation lysis buffer/product/Millipore
    Average 86 stars, based on 1 article reviews
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    92
    Millipore rip lysis buffer
    HK2 is a target of miR-125a. a The putative binding sites of miR-125a on the 3′ UTR of HK2. b The protein levels of HK2 in <t>AMC-HN-8</t> and TU212 cells 48 h after miR-125a transfection. c Relative activity of luciferase reporters with HK2 3′ UTR after co-transfection with miR-125a mimics in AMC-HN-8 and TU212 cells. d Cellular lysates from AMC-HN-8 and TU212 cells were used for RNA immunoprecipitation <t>(RIP)</t> with Ago2 antibody. miR-125a and HK2 mRNA were detected using qRT-PCR. * P
    Rip Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rip lysis buffer/product/Millipore
    Average 92 stars, based on 98 article reviews
    Price from $9.99 to $1999.99
    rip lysis buffer - by Bioz Stars, 2020-09
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    85
    Millipore empigen containing cell lysis buffer
    Endogenous hAgo2 and hAgo3 Slicer activities. ( A ) Immunopurified hAgo2 from HeLa cells, which contained miR-21 (shown in B ), subjected to in vitro target RNA cleavage assays. Even hAgo2 immunopurified from HeLa cells in an <t>Empigen</t> buffer shows Slicer
    Empigen Containing Cell Lysis Buffer, supplied by Millipore, 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/empigen containing cell lysis buffer/product/Millipore
    Average 85 stars, based on 1 article reviews
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    empigen containing cell lysis buffer - by Bioz Stars, 2020-09
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    CircCLK3 expression is relatively highly expressed in cervical cancer tissues and predicts poor prognosis. a CircRNA sequencing was adapted to detect the differentially expressed circRNAs between cervical cancer tissues and matched normal tissues. b The genomic loci of the CLK3 gene and circCLK3. Sanger sequencing confirmed the head-to-hail splicing. c The expression level of circCLK3 in 5 cervical cancer cell lines relative to HeLa cell. d The gel electrophoresis validated the existence of circCLK3. Divergent primers amplified circCLK3 in cDNA but not gDNA. e qRT-PCR results of U6, GAPDH, and circCLK3 expressions in cell nuclei and cytoplasm in SiHa and HeLa cells. f RNA-FISH assays were performed to identify the subcellular location of circCLK3 with the cy3-labeled circCLK3 probe. The results showed that most of circCLK3 was located in cytoplasm in SiHa and HeLa cells. g qRT-PCR results of circCLK3 and CLK3 expression after treatment with actinomycin D at the indicated time points in SiHa and HeLa cells. h qRT-PCR results of circCLK3 and CLK3 expression after treatment with RNase R in SiHa and HeLa cells. i CircCLK3 expression was significantly higher in 48 paired fresh frozen cervical cancer tissues compared with adjacent normal tissues. j , k Kaplan–Meier Plotter analysis of the correlation between circCLK3 level with overall survive or disease-free survive of cervical cancer patients. Data are reported as means ± standard deviation of three independent experiments. * p

    Journal: Cell Death & Disease

    Article Title: The novel circCLK3/miR-320a/FoxM1 axis promotes cervical cancer progression

    doi: 10.1038/s41419-019-2183-z

    Figure Lengend Snippet: CircCLK3 expression is relatively highly expressed in cervical cancer tissues and predicts poor prognosis. a CircRNA sequencing was adapted to detect the differentially expressed circRNAs between cervical cancer tissues and matched normal tissues. b The genomic loci of the CLK3 gene and circCLK3. Sanger sequencing confirmed the head-to-hail splicing. c The expression level of circCLK3 in 5 cervical cancer cell lines relative to HeLa cell. d The gel electrophoresis validated the existence of circCLK3. Divergent primers amplified circCLK3 in cDNA but not gDNA. e qRT-PCR results of U6, GAPDH, and circCLK3 expressions in cell nuclei and cytoplasm in SiHa and HeLa cells. f RNA-FISH assays were performed to identify the subcellular location of circCLK3 with the cy3-labeled circCLK3 probe. The results showed that most of circCLK3 was located in cytoplasm in SiHa and HeLa cells. g qRT-PCR results of circCLK3 and CLK3 expression after treatment with actinomycin D at the indicated time points in SiHa and HeLa cells. h qRT-PCR results of circCLK3 and CLK3 expression after treatment with RNase R in SiHa and HeLa cells. i CircCLK3 expression was significantly higher in 48 paired fresh frozen cervical cancer tissues compared with adjacent normal tissues. j , k Kaplan–Meier Plotter analysis of the correlation between circCLK3 level with overall survive or disease-free survive of cervical cancer patients. Data are reported as means ± standard deviation of three independent experiments. * p

    Article Snippet: SiHa cells were transfected with miR-320a mimics, and then lysed in complete RNA lysis buffer after 48 h. Cell lysates were rotated in RIP immunoprecipitation buffer including magnetic beads conjugated with negative control mouse IgG or human anti-AGO2 antibody (Mouse, Millipore, Billerica, USA) overnight.

    Techniques: Expressing, Sequencing, Nucleic Acid Electrophoresis, Amplification, Quantitative RT-PCR, Fluorescence In Situ Hybridization, Labeling, Standard Deviation

    PIWIL1 cooperates with UPF1 to negatively regulate PIWIL1-bound RNAs in a piRNA-independent manner. (A) A Coomassie blue staining protein gel of PIWIL1-co-immunoprecipitation, Mass spectrometry of this protein gel identifies PIWIL1 (blue arrow) as well as UPF1 (red arrow). (B) Western blotting showing reciprocal co-immunoprecipitated between PIWIL1 and UPF1 (total and phosphorylated form, p-UPF1) and between PIWIL1 and UPF2. (C) Western blotting shows that PIWIL1 co-immunoprecipitates with NMD complex core proteins UPF1, phosphorylated UPF1 (p-UPF1), UPF2, and SMG1. (D) Immunofluorescence staining of PIWIL1 (green) and DCP1A (red, a P body marker) in wildtype and PIWIL_KO SNU-1 cells. (E) Immunofluorescence staining of PIWIL1 (green), DCP1A (red), and UPF1 (fuchsia) in SNU-1 cells, which shows PIWIL1 co-localized with UPF1 in the P body. (F) Venn diagram of PIWIL1-bound RNAs, UPF1-bound RNAs, and PIWIL1-negatively regulated RNAs, with P

    Journal: bioRxiv

    Article Title: PIWIL1 Promotes Gastric Cancer via a piRNA-Independent Mechanism

    doi: 10.1101/2020.05.03.075390

    Figure Lengend Snippet: PIWIL1 cooperates with UPF1 to negatively regulate PIWIL1-bound RNAs in a piRNA-independent manner. (A) A Coomassie blue staining protein gel of PIWIL1-co-immunoprecipitation, Mass spectrometry of this protein gel identifies PIWIL1 (blue arrow) as well as UPF1 (red arrow). (B) Western blotting showing reciprocal co-immunoprecipitated between PIWIL1 and UPF1 (total and phosphorylated form, p-UPF1) and between PIWIL1 and UPF2. (C) Western blotting shows that PIWIL1 co-immunoprecipitates with NMD complex core proteins UPF1, phosphorylated UPF1 (p-UPF1), UPF2, and SMG1. (D) Immunofluorescence staining of PIWIL1 (green) and DCP1A (red, a P body marker) in wildtype and PIWIL_KO SNU-1 cells. (E) Immunofluorescence staining of PIWIL1 (green), DCP1A (red), and UPF1 (fuchsia) in SNU-1 cells, which shows PIWIL1 co-localized with UPF1 in the P body. (F) Venn diagram of PIWIL1-bound RNAs, UPF1-bound RNAs, and PIWIL1-negatively regulated RNAs, with P

    Article Snippet: Co-immunoprecipitation Cells were lysed in co-immunoprecipitation lysis buffer [20mM Tris-HCl pH7.4, 150mM NaCl, 1 % (v/v) IGEPAL® CA-630 (Millipore SIGMA, Cat# I8896), 1mM EDTA, 0.5 mM DTT, cOmplete™ EDTA-free Protease Inhibitor Cocktail Tablets (MERCK, Cat# 4693132001), and PhosStop Tablets (MERCK, 4906837001)], and spun at 14,000 rpm for 10 minutes to remove the debris.

    Techniques: Staining, Immunoprecipitation, Mass Spectrometry, Western Blot, Immunofluorescence, Marker

    HK2 is a target of miR-125a. a The putative binding sites of miR-125a on the 3′ UTR of HK2. b The protein levels of HK2 in AMC-HN-8 and TU212 cells 48 h after miR-125a transfection. c Relative activity of luciferase reporters with HK2 3′ UTR after co-transfection with miR-125a mimics in AMC-HN-8 and TU212 cells. d Cellular lysates from AMC-HN-8 and TU212 cells were used for RNA immunoprecipitation (RIP) with Ago2 antibody. miR-125a and HK2 mRNA were detected using qRT-PCR. * P

    Journal: Cell & Bioscience

    Article Title: miR-125a suppresses viability and glycolysis and induces apoptosis by targeting Hexokinase 2 in laryngeal squamous cell carcinoma

    doi: 10.1186/s13578-017-0178-y

    Figure Lengend Snippet: HK2 is a target of miR-125a. a The putative binding sites of miR-125a on the 3′ UTR of HK2. b The protein levels of HK2 in AMC-HN-8 and TU212 cells 48 h after miR-125a transfection. c Relative activity of luciferase reporters with HK2 3′ UTR after co-transfection with miR-125a mimics in AMC-HN-8 and TU212 cells. d Cellular lysates from AMC-HN-8 and TU212 cells were used for RNA immunoprecipitation (RIP) with Ago2 antibody. miR-125a and HK2 mRNA were detected using qRT-PCR. * P

    Article Snippet: Briefly, AMC-HN-8 and TU212 cells were lysed in RIP lysis buffer, then 100 μl of whole cell extract was incubated with RIP buffer containing magnetic beads conjugated with human anti-Ago2 antibody, positive control anti-snRNP70 or negative control normal mouse IgG (Millipore).

    Techniques: Binding Assay, Transfection, Activity Assay, Luciferase, Cotransfection, Immunoprecipitation, Quantitative RT-PCR

    Endogenous hAgo2 and hAgo3 Slicer activities. ( A ) Immunopurified hAgo2 from HeLa cells, which contained miR-21 (shown in B ), subjected to in vitro target RNA cleavage assays. Even hAgo2 immunopurified from HeLa cells in an Empigen buffer shows Slicer

    Journal:

    Article Title: Characterization of endogenous human Argonautes and their miRNA partners in RNA silencing

    doi: 10.1073/pnas.0800334105

    Figure Lengend Snippet: Endogenous hAgo2 and hAgo3 Slicer activities. ( A ) Immunopurified hAgo2 from HeLa cells, which contained miR-21 (shown in B ), subjected to in vitro target RNA cleavage assays. Even hAgo2 immunopurified from HeLa cells in an Empigen buffer shows Slicer

    Article Snippet: Empigen-containing cell lysis buffer was 1× PBS containing 1 mM EDTA, 0.1 mM DTT, 1% Empigen (Calbiochem), 2 μg/ml pepstatin, 2 μg/ml leupeptin, and 0.5% aprotinin.

    Techniques: In Vitro

    Characterization of immunoprecipitates with anti-hAgo2, -hAgo3, and -hAgo4 antibodies from Jurkat cells. ( A ) Jurkat cell lysates prepared with a cell lysis buffer containing Empigen, and immunoprecipitation performed by using anti-hAgo2, -hAgo3, and -hAgo4

    Journal:

    Article Title: Characterization of endogenous human Argonautes and their miRNA partners in RNA silencing

    doi: 10.1073/pnas.0800334105

    Figure Lengend Snippet: Characterization of immunoprecipitates with anti-hAgo2, -hAgo3, and -hAgo4 antibodies from Jurkat cells. ( A ) Jurkat cell lysates prepared with a cell lysis buffer containing Empigen, and immunoprecipitation performed by using anti-hAgo2, -hAgo3, and -hAgo4

    Article Snippet: Empigen-containing cell lysis buffer was 1× PBS containing 1 mM EDTA, 0.1 mM DTT, 1% Empigen (Calbiochem), 2 μg/ml pepstatin, 2 μg/ml leupeptin, and 0.5% aprotinin.

    Techniques: Lysis, Immunoprecipitation