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  • 93
    Thermo Fisher superscriptii platinum taq long range one step rt pcr kit
    Superscriptii Platinum Taq Long Range One Step Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen long range rt pcr kit
    Characterization of ectopically expressed RNAs by long range <t>RT-PCR.</t> MDA-MB-231 parental cell line (231-parental), its pcDNA3-ERα stable transfectant (ERα-2), and its ERα-IRES stable transfectants (ERα-IRES-5 and ERα-IRES-3) were analyzed for expression of ERα–harboring transcript (1.8 kb), Hygromycin B resistance gene-containing transcript (1.0 kb), and ERα-IRES-Hygro R fused transcript (3.2 kb), by RT followed by long range PCR amplification. pERα-IRES DNA served as a PCR positive control for the ERα <t>cDNA</t> primers (1.8 kb), the Hygromycin B resistance gene ORF primers (1.0 kb), and the 5′ sense ERα primer plus 3′ antisense Hygro R fused ORFs primers (3.5 kb). A First four lanes from left contain the ERα cDNA primers; lanes 5–8 the 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. B. The 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. C. Lanes 1 and 2 from left, the ERα primers. Lanes 3 and 4 the Hygro R gene primers. Primer sequences are detailed in the “ Methods ” section.
    Long Range Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eppendorf AG long range rt pcr kit
    Characterization of ectopically expressed RNAs by long range <t>RT-PCR.</t> MDA-MB-231 parental cell line (231-parental), its pcDNA3-ERα stable transfectant (ERα-2), and its ERα-IRES stable transfectants (ERα-IRES-5 and ERα-IRES-3) were analyzed for expression of ERα–harboring transcript (1.8 kb), Hygromycin B resistance gene-containing transcript (1.0 kb), and ERα-IRES-Hygro R fused transcript (3.2 kb), by RT followed by long range PCR amplification. pERα-IRES DNA served as a PCR positive control for the ERα <t>cDNA</t> primers (1.8 kb), the Hygromycin B resistance gene ORF primers (1.0 kb), and the 5′ sense ERα primer plus 3′ antisense Hygro R fused ORFs primers (3.5 kb). A First four lanes from left contain the ERα cDNA primers; lanes 5–8 the 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. B. The 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. C. Lanes 1 and 2 from left, the ERα primers. Lanes 3 and 4 the Hygro R gene primers. Primer sequences are detailed in the “ Methods ” section.
    Long Range Rt Pcr Kit, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher one tube long range rt pcr kit
    Characterization of ectopically expressed RNAs by long range <t>RT-PCR.</t> MDA-MB-231 parental cell line (231-parental), its pcDNA3-ERα stable transfectant (ERα-2), and its ERα-IRES stable transfectants (ERα-IRES-5 and ERα-IRES-3) were analyzed for expression of ERα–harboring transcript (1.8 kb), Hygromycin B resistance gene-containing transcript (1.0 kb), and ERα-IRES-Hygro R fused transcript (3.2 kb), by RT followed by long range PCR amplification. pERα-IRES DNA served as a PCR positive control for the ERα <t>cDNA</t> primers (1.8 kb), the Hygromycin B resistance gene ORF primers (1.0 kb), and the 5′ sense ERα primer plus 3′ antisense Hygro R fused ORFs primers (3.5 kb). A First four lanes from left contain the ERα cDNA primers; lanes 5–8 the 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. B. The 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. C. Lanes 1 and 2 from left, the ERα primers. Lanes 3 and 4 the Hygro R gene primers. Primer sequences are detailed in the “ Methods ” section.
    One Tube Long Range Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche long range pcr kit
    IL-1β induction and chromatin structure of cPLA 2 <t>α</t> in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time <t>RT-PCR.</t> Relative fold induction was calculated
    Long Range Pcr Kit, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen one step rt pcr kit
    IL-1β induction and chromatin structure of cPLA 2 <t>α</t> in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time <t>RT-PCR.</t> Relative fold induction was calculated
    One Step Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 12631 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa exscript rt pcr kit
    <t>STC2</t> maintains breast cancer cell morphology. A , Analyses of STC2 by Western blotting and <t>realtime-PCR</t> in constructed cells (P
    Exscript Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 762 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of ectopically expressed RNAs by long range RT-PCR. MDA-MB-231 parental cell line (231-parental), its pcDNA3-ERα stable transfectant (ERα-2), and its ERα-IRES stable transfectants (ERα-IRES-5 and ERα-IRES-3) were analyzed for expression of ERα–harboring transcript (1.8 kb), Hygromycin B resistance gene-containing transcript (1.0 kb), and ERα-IRES-Hygro R fused transcript (3.2 kb), by RT followed by long range PCR amplification. pERα-IRES DNA served as a PCR positive control for the ERα cDNA primers (1.8 kb), the Hygromycin B resistance gene ORF primers (1.0 kb), and the 5′ sense ERα primer plus 3′ antisense Hygro R fused ORFs primers (3.5 kb). A First four lanes from left contain the ERα cDNA primers; lanes 5–8 the 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. B. The 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. C. Lanes 1 and 2 from left, the ERα primers. Lanes 3 and 4 the Hygro R gene primers. Primer sequences are detailed in the “ Methods ” section.

    Journal: PLoS ONE

    Article Title: ER-Alpha-cDNA As Part of a Bicistronic Transcript Gives Rise to High Frequency, Long Term, Receptor Expressing Cell Clones

    doi: 10.1371/journal.pone.0031977

    Figure Lengend Snippet: Characterization of ectopically expressed RNAs by long range RT-PCR. MDA-MB-231 parental cell line (231-parental), its pcDNA3-ERα stable transfectant (ERα-2), and its ERα-IRES stable transfectants (ERα-IRES-5 and ERα-IRES-3) were analyzed for expression of ERα–harboring transcript (1.8 kb), Hygromycin B resistance gene-containing transcript (1.0 kb), and ERα-IRES-Hygro R fused transcript (3.2 kb), by RT followed by long range PCR amplification. pERα-IRES DNA served as a PCR positive control for the ERα cDNA primers (1.8 kb), the Hygromycin B resistance gene ORF primers (1.0 kb), and the 5′ sense ERα primer plus 3′ antisense Hygro R fused ORFs primers (3.5 kb). A First four lanes from left contain the ERα cDNA primers; lanes 5–8 the 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. B. The 5′ sense ERα primer together with the 3′ antisense Hygro R gene primer. C. Lanes 1 and 2 from left, the ERα primers. Lanes 3 and 4 the Hygro R gene primers. Primer sequences are detailed in the “ Methods ” section.

    Article Snippet: Two µg of total RNA extracted using EZ-RNA isolation kit (Biological Industries, Israel) were transcribed into first strand cDNA by hexamer priming, followed by PCR reactions as specified in the Long range RT-PCR kit (Qiagen).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Multiple Displacement Amplification, Transfection, Expressing, Polymerase Chain Reaction, Amplification, Positive Control

    IL-1β induction and chromatin structure of cPLA 2 α in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time RT-PCR. Relative fold induction was calculated

    Journal: Journal of Lipid Research

    Article Title: A distal enhancer controls cytokine-dependent human cPLA2? gene expression [S]

    doi: 10.1194/jlr.M037382

    Figure Lengend Snippet: IL-1β induction and chromatin structure of cPLA 2 α in HFL-1. (A, left) HFL-1 cells were treated with 2 ng/µl IL-1β for the indicated times, and total RNA was analyzed by real-time RT-PCR. Relative fold induction was calculated

    Article Snippet: The −14, −5.5, and −4.1 kb cPLA2 α promoters were amplified from using the Long Range PCR kit (Roche), with the forward primers 5′-AGAGTTGGGATGGAGAAGGTTG-3′, 5′-TGCAAAGTGCCTGCCAGTC-3′, or 5′-GATGGAGATGGCAGTGGCAG-3′, respectively, and the reverse primer 5′-GCTTACAGTTCCCAGAGTTACC-3′, and cloned into the TOPO-XL vector.

    Techniques: Quantitative RT-PCR

    STC2 maintains breast cancer cell morphology. A , Analyses of STC2 by Western blotting and realtime-PCR in constructed cells (P

    Journal: PLoS ONE

    Article Title: Stanniocalicin 2 Suppresses Breast Cancer Cell Migration and Invasion via the PKC/Claudin-1-Mediated Signaling

    doi: 10.1371/journal.pone.0122179

    Figure Lengend Snippet: STC2 maintains breast cancer cell morphology. A , Analyses of STC2 by Western blotting and realtime-PCR in constructed cells (P

    Article Snippet: Real-time PCR Total RNA was isolated from 2 × 106 231 STC2 or HM STC2i cells, and their control cells, by using Trizol reagent (Invitrogen, Carlsbad, CA). cDNAs generated by reverse transcription were used for real-time PCR analysis with the ExScript RT-PCR kit (TaKaRa, Japan). qPCR primers for STC2 are as follows: 5’-GGGTGTGGCGTGTTTGAATG-3’ (sense) and 5’-CTTGAGGTAGCATTCCCGCT-3’ (antisense). qPCR primers for GAPDH are as follows: 5’- ACCCAGAAGACTGTGGATGG-3’ (sense) and 5’-TCTAGACGGCAGGTCAGGTC-3’ (antisense).

    Techniques: Western Blot, Polymerase Chain Reaction, Construct

    Effects of rosuvastatin on electrical remodeling after acute MI. The mRNA expression level of KCND3 potassium ion channel was detected by real-time PCR in atrial tissues following MI, in the sham, MI model, and intervention groups. Compared with the sham

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Effects of rosuvastatin on atrial nerve sprouting and electrical remodeling in rabbits with myocardial infarction

    doi:

    Figure Lengend Snippet: Effects of rosuvastatin on electrical remodeling after acute MI. The mRNA expression level of KCND3 potassium ion channel was detected by real-time PCR in atrial tissues following MI, in the sham, MI model, and intervention groups. Compared with the sham

    Article Snippet: The mRNA expression levels of TH and KCND3were detected with the RT-PCR reagent kit (Takara, Dalian, Liaoning, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Effects of rosuvastatin on TH expression level in atrial tissues after acute MI. The mRNA and protein expression levels of TH in rabbit atrial tissues were detected by real-Time PCR (A) and Western blot analysis (B), respectively. Compared with the sham

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Effects of rosuvastatin on atrial nerve sprouting and electrical remodeling in rabbits with myocardial infarction

    doi:

    Figure Lengend Snippet: Effects of rosuvastatin on TH expression level in atrial tissues after acute MI. The mRNA and protein expression levels of TH in rabbit atrial tissues were detected by real-Time PCR (A) and Western blot analysis (B), respectively. Compared with the sham

    Article Snippet: The mRNA expression levels of TH and KCND3were detected with the RT-PCR reagent kit (Takara, Dalian, Liaoning, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    Expression of cell surface molecules in lymph node-derived stromal cells. (a) Stromal cells derived from lymph nodes of IL-2Rβ −/− mice were stained with fluorescence-conjugated antibodies and were analysed by flow cytometry. Shaded peaks and open peaks with dotted lines indicate the fluorescence intensities of cells stained with a non-specific control antibody labelled with the same fluorescent reagents. (b) RNA was extracted from stromal cells derived from lymph nodes of IL-2Rβ −/− mice, and RT–PCR was performed using primers to detect the expression of indicated cytokines.

    Journal: Immunology

    Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro

    doi: 10.1046/j.1365-2567.2003.01693.x

    Figure Lengend Snippet: Expression of cell surface molecules in lymph node-derived stromal cells. (a) Stromal cells derived from lymph nodes of IL-2Rβ −/− mice were stained with fluorescence-conjugated antibodies and were analysed by flow cytometry. Shaded peaks and open peaks with dotted lines indicate the fluorescence intensities of cells stained with a non-specific control antibody labelled with the same fluorescent reagents. (b) RNA was extracted from stromal cells derived from lymph nodes of IL-2Rβ −/− mice, and RT–PCR was performed using primers to detect the expression of indicated cytokines.

    Article Snippet: Single-stranded cDNA was synthesized by reverse transcriptase (RT) with random hexamer oligonucleotides as primers, and the desired cDNA was amplified by polymerase chain reaction (PCR) using a TaKaRa RNA LA PCR kit (TaKaRa Biomedicals, Kyoto, Japan).

    Techniques: Expressing, Derivative Assay, Mouse Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Reverse Transcription Polymerase Chain Reaction

    Breakpoint determination. a PALB2 exon 13 duplication confirmation by long-range PCR. An extra band of approximately 4–6 kb in size was detected in the proband’s blood DNA, which is absent in the negative control sample. b Localization of breakpoint. Electropherogram showing the breakpoint sequence. c Sequence alignment in the breakpoint region showing the 32-bp perfect identity between the intron 12 and 3′-UTR of PALB2

    Journal: Breast cancer research and treatment

    Article Title: Characterization of a novel germline PALB2 duplication in a hereditary breast and ovarian cancer family

    doi: 10.1007/s10549-016-4021-7

    Figure Lengend Snippet: Breakpoint determination. a PALB2 exon 13 duplication confirmation by long-range PCR. An extra band of approximately 4–6 kb in size was detected in the proband’s blood DNA, which is absent in the negative control sample. b Localization of breakpoint. Electropherogram showing the breakpoint sequence. c Sequence alignment in the breakpoint region showing the 32-bp perfect identity between the intron 12 and 3′-UTR of PALB2

    Article Snippet: LR-PCR was performed using a forward primer in intron 12 and a reverse primer in 3′-untranslated region (UTR) of PALB2 and the TaKaRa LA PCR kit (TaKaRa, Clontech) following the manufacturer’s instructions.

    Techniques: Polymerase Chain Reaction, Negative Control, Sequencing

    An Alu-mediated mechanism appears to be responsible for exon 13 duplication. A sequence analysis of the LR-PCR product allowed us to identify the duplication conjunctions which are located 1089 bp downstream of exon 12, in intron 12, and 989 bp downstream of the PALB2 3′-UTR

    Journal: Breast cancer research and treatment

    Article Title: Characterization of a novel germline PALB2 duplication in a hereditary breast and ovarian cancer family

    doi: 10.1007/s10549-016-4021-7

    Figure Lengend Snippet: An Alu-mediated mechanism appears to be responsible for exon 13 duplication. A sequence analysis of the LR-PCR product allowed us to identify the duplication conjunctions which are located 1089 bp downstream of exon 12, in intron 12, and 989 bp downstream of the PALB2 3′-UTR

    Article Snippet: LR-PCR was performed using a forward primer in intron 12 and a reverse primer in 3′-untranslated region (UTR) of PALB2 and the TaKaRa LA PCR kit (TaKaRa, Clontech) following the manufacturer’s instructions.

    Techniques: Sequencing, Polymerase Chain Reaction

    PALB2 Exon 13 duplication disrupts normal splicing and leads to frameshift. a RT-PCR products run on QIAxcel. An extra band was observed in the patient, patient’s 48-year-old sister, aunt, and 52-year-old sister. b Electropherogram showing the inserted sequence from the upper band in the patient of figure ( a ). The insertion leads to a new stop codon as indicated by the red box

    Journal: Breast cancer research and treatment

    Article Title: Characterization of a novel germline PALB2 duplication in a hereditary breast and ovarian cancer family

    doi: 10.1007/s10549-016-4021-7

    Figure Lengend Snippet: PALB2 Exon 13 duplication disrupts normal splicing and leads to frameshift. a RT-PCR products run on QIAxcel. An extra band was observed in the patient, patient’s 48-year-old sister, aunt, and 52-year-old sister. b Electropherogram showing the inserted sequence from the upper band in the patient of figure ( a ). The insertion leads to a new stop codon as indicated by the red box

    Article Snippet: LR-PCR was performed using a forward primer in intron 12 and a reverse primer in 3′-untranslated region (UTR) of PALB2 and the TaKaRa LA PCR kit (TaKaRa, Clontech) following the manufacturer’s instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Sequencing

    Activation of NFκB-response element in the 2 nd intron of MnSOD gene by BTG2. (A) MnSOD promoter analysis; Cells (0.5×10 5 /12 wells) were cotransfected with BTG2 cDNA and promoter DNA of MnSOD gene and then subjected to luciferase analysis. Transfection of BTG2 up to 100 ng failed to activate promoter of MnSOD. Sp1 cDNA was employed as a positive control. Lower panel shows protein expressions of the transfected DNAs and loading control. (B) Cells were cotransfected with BTG2 cDNA and the κB-RE before luciferase assay. Expression of MnSOD was significantly increased with transfection of BTG2, indicating the activation of κB-RE by BTG2. Immunoblot analysis shows protein expression of BTG2-HA and loading control. (C) To further investigate the regulation of NFκB activation and MnSOD induction by BTG2 expression, IκBα degradation was examined by immunoblot analysis. Note the degradation of IκBα and MnSOD expression in the BTG2-dependent manner. (D) Transduction of HeLa cells with Ad-BTG2 was performed and then treated with 10 μM of MG132 for 1 h before RT-PCR analysis. MG132 abolished induction of MnSOD expression by BTG2, suggesting the upregulation of MnSOD expression by BTG2 via proteasomal degradation of IκBα. (E) Cell lysates with the LacZ or the BTG2 expressers were subjected to ChIP analysis with anti-p65 antibody, and then the interaction was verified by PCR reaction with the primers written in the Additional file 5 . To verify our analysis, HeLa cells were treated with or without 100 ng of 12- O -tetradecanoylphorbol-13-acetate (TPA) for 2 h and then applied to ChIP assay as a positive and negative control. Unstimulated IgG was employed to exclude the nonspecific interaction. Inputs indicate total amount of κB-RE present in the samples. Note the interactions of p65 with κB-RE only in the BTG2 expressers and the TPA treated positive cells. (F) Immunoblot analyses showing the activation of IKKα/β in the BTG2 overexpressers without any changes in the expression of other anti-oxidant enzymes.

    Journal: Cell Communication and Signaling : CCS

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells

    doi: 10.1186/1478-811X-11-69

    Figure Lengend Snippet: Activation of NFκB-response element in the 2 nd intron of MnSOD gene by BTG2. (A) MnSOD promoter analysis; Cells (0.5×10 5 /12 wells) were cotransfected with BTG2 cDNA and promoter DNA of MnSOD gene and then subjected to luciferase analysis. Transfection of BTG2 up to 100 ng failed to activate promoter of MnSOD. Sp1 cDNA was employed as a positive control. Lower panel shows protein expressions of the transfected DNAs and loading control. (B) Cells were cotransfected with BTG2 cDNA and the κB-RE before luciferase assay. Expression of MnSOD was significantly increased with transfection of BTG2, indicating the activation of κB-RE by BTG2. Immunoblot analysis shows protein expression of BTG2-HA and loading control. (C) To further investigate the regulation of NFκB activation and MnSOD induction by BTG2 expression, IκBα degradation was examined by immunoblot analysis. Note the degradation of IκBα and MnSOD expression in the BTG2-dependent manner. (D) Transduction of HeLa cells with Ad-BTG2 was performed and then treated with 10 μM of MG132 for 1 h before RT-PCR analysis. MG132 abolished induction of MnSOD expression by BTG2, suggesting the upregulation of MnSOD expression by BTG2 via proteasomal degradation of IκBα. (E) Cell lysates with the LacZ or the BTG2 expressers were subjected to ChIP analysis with anti-p65 antibody, and then the interaction was verified by PCR reaction with the primers written in the Additional file 5 . To verify our analysis, HeLa cells were treated with or without 100 ng of 12- O -tetradecanoylphorbol-13-acetate (TPA) for 2 h and then applied to ChIP assay as a positive and negative control. Unstimulated IgG was employed to exclude the nonspecific interaction. Inputs indicate total amount of κB-RE present in the samples. Note the interactions of p65 with κB-RE only in the BTG2 expressers and the TPA treated positive cells. (F) Immunoblot analyses showing the activation of IKKα/β in the BTG2 overexpressers without any changes in the expression of other anti-oxidant enzymes.

    Article Snippet: RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume.

    Techniques: Activation Assay, Luciferase, Transfection, Positive Control, Expressing, Transduction, Reverse Transcription Polymerase Chain Reaction, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Negative Control