Structured Review

Roche genomic pcr
Monoallelic assembly of <t>VLRA</t> and VLRB genes a,b, Schematic of VLRA ( a ) and VLRB ( b ) genes before (top panel) and after (middle panel) gene assembly. Forward and reverse primer locations and predicted sizes of <t>PCR</t> products are indicated. Lymphocytes from the indicated tissues were stained with anti-VLRA (R110) and anti-VLR-B (4C4) antibodies and lymphocyte-gated cells were FACS sorted into three populations: VLRA − /VLRB − (DN), VLRA + (A + ), and VLRB + (B + ). VLRs were amplified from genomic DNA of the sorted lymphocyte populations (bottom panel). Germ-line (GL) and mature (M) products were verified by sequence analysis of representative DNA clones. c , CDA1 and CDA2 expression in sorted lymphocytes was measured by QPCR. Error bars indicate s.e.m., n = 3.
Genomic Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Dual Nature of the Adaptive Immune System in Lampreys"

Article Title: Dual Nature of the Adaptive Immune System in Lampreys

Journal: Nature

doi: 10.1038/nature08068

Monoallelic assembly of VLRA and VLRB genes a,b, Schematic of VLRA ( a ) and VLRB ( b ) genes before (top panel) and after (middle panel) gene assembly. Forward and reverse primer locations and predicted sizes of PCR products are indicated. Lymphocytes from the indicated tissues were stained with anti-VLRA (R110) and anti-VLR-B (4C4) antibodies and lymphocyte-gated cells were FACS sorted into three populations: VLRA − /VLRB − (DN), VLRA + (A + ), and VLRB + (B + ). VLRs were amplified from genomic DNA of the sorted lymphocyte populations (bottom panel). Germ-line (GL) and mature (M) products were verified by sequence analysis of representative DNA clones. c , CDA1 and CDA2 expression in sorted lymphocytes was measured by QPCR. Error bars indicate s.e.m., n = 3.
Figure Legend Snippet: Monoallelic assembly of VLRA and VLRB genes a,b, Schematic of VLRA ( a ) and VLRB ( b ) genes before (top panel) and after (middle panel) gene assembly. Forward and reverse primer locations and predicted sizes of PCR products are indicated. Lymphocytes from the indicated tissues were stained with anti-VLRA (R110) and anti-VLR-B (4C4) antibodies and lymphocyte-gated cells were FACS sorted into three populations: VLRA − /VLRB − (DN), VLRA + (A + ), and VLRB + (B + ). VLRs were amplified from genomic DNA of the sorted lymphocyte populations (bottom panel). Germ-line (GL) and mature (M) products were verified by sequence analysis of representative DNA clones. c , CDA1 and CDA2 expression in sorted lymphocytes was measured by QPCR. Error bars indicate s.e.m., n = 3.

Techniques Used: Polymerase Chain Reaction, Staining, FACS, Amplification, Sequencing, Clone Assay, Expressing, Real-time Polymerase Chain Reaction

Related Articles

Polymerase Chain Reaction:

Article Title: Recurrent ESR1-CCDC170 rearrangements in an aggressive subset of estrogen-receptor positive breast cancers
Article Snippet: .. Genomic PCR was carried out with 200-300ng of genomic DNA from cell lines or tissues using the Expand Long Range PCR system (Roche) and primers listed in . ..

Article Title: Dual Nature of the Adaptive Immune System in Lampreys
Article Snippet: .. Genomic PCR was carried out using primers VLRA-F + VLRA-R (Expand High Fidelity, Roche and Ex Taq, Takara) or VLRB-F + VLRB-R (Expand Long Template, Roche). .. Quantitative PCR Target gene sequences were obtained from the National Center for Biotechnology Information database or the lamprey genome database of the Genome Sequencing Center at Washington University.

Article Title: Evolutionary history of teleost intron-containing and intron-less rhodopsin genes
Article Snippet: .. Genomic PCR and RT-PCR analysis Genomic DNA was isolated from gray bichir, reedfish and Siberian sturgeon using High Pure PCR Template Preparation Kit (Roche). .. Total RNA was isolated from eyes of gray bichir, reedfish and Siberian sturgeon using RNeasy Plus Mini Kit (QIAGEN).

Article Title: Evolutionary implications of a third lymphocyte lineage in lampreys
Article Snippet: .. Genomic PCR was carried out using primers VLRA-F and VLRA-R, VLRB-F and VLRB-R or VLRC-F and VLRC-R (Expand Long Template, Roche). .. Total RNA was extracted from sorted cells of lamprey blood and tissues (kidneys, typhlosole, gill and skin) and reverse-transcribed using random hexamers (Invitrogen).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Evolutionary history of teleost intron-containing and intron-less rhodopsin genes
Article Snippet: .. Genomic PCR and RT-PCR analysis Genomic DNA was isolated from gray bichir, reedfish and Siberian sturgeon using High Pure PCR Template Preparation Kit (Roche). .. Total RNA was isolated from eyes of gray bichir, reedfish and Siberian sturgeon using RNeasy Plus Mini Kit (QIAGEN).

Isolation:

Article Title: Evolutionary history of teleost intron-containing and intron-less rhodopsin genes
Article Snippet: .. Genomic PCR and RT-PCR analysis Genomic DNA was isolated from gray bichir, reedfish and Siberian sturgeon using High Pure PCR Template Preparation Kit (Roche). .. Total RNA was isolated from eyes of gray bichir, reedfish and Siberian sturgeon using RNeasy Plus Mini Kit (QIAGEN).

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    STAT3 binding to promoter region hTERT were inhibited by the resveratrol and 5-FU alone and in combination and corresponding <t>telomerase</t> activities were decreased upon the combination treatments Transcription factor STAT3 binding to hTERT promoter region was tested with chromatin immunoprecipitation assay. (A) HCT116 was treated with 5-FU and resveratrol alone and in combination and applied to ChIP assay. STAT3 binding was quantified by the ChIP band intensity relative to untreated control, which was set as 1.0. (B) Telomerase activities were measured by TRAP PCR reaction conjugated to Elisa assay. (C) DLD1 was treated with 5-FU (10 μM) alone and associated with 25 μM resveratrol combination, then applied to ChIP assay to quantify the STAT3 binding. (D) DLD1 was treated with 5-FU and resveratrol alone and in combination and applied to TRAP-PCR-Elisa assay to quantify telomerase activities. Error bars represent standard deviation. Telomerase activities were measured three times independently.
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    Antibody competition titration assays using MSP1 19 proteins from four Plasmodium species. A combined dilution (1:400 of each serum) containing sera from chimpanzees experimentally infected with either P. <t>malariae</t> (Klimatis), P. ovale (Alpert) or P. vivax (Duff) was incubated with the indicated concentrations of the MSP1 19 competitor protein for 1 h at room temperature. Competitor proteins used were: a P. falciparum MSP1 19 ; b P. malariae MSP1 19 ; c P. ovale MSP1 19 ; d P. vivax MSP1 19 . Multiplex bead assays were performed as described in “ Methods ” and the multiplex response in MFI-bg units are plotted versus the competitor concentration. Multiplex responses are presented as a percentage of the assay results for the PBS control
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    STAT3 binding to promoter region hTERT were inhibited by the resveratrol and 5-FU alone and in combination and corresponding telomerase activities were decreased upon the combination treatments Transcription factor STAT3 binding to hTERT promoter region was tested with chromatin immunoprecipitation assay. (A) HCT116 was treated with 5-FU and resveratrol alone and in combination and applied to ChIP assay. STAT3 binding was quantified by the ChIP band intensity relative to untreated control, which was set as 1.0. (B) Telomerase activities were measured by TRAP PCR reaction conjugated to Elisa assay. (C) DLD1 was treated with 5-FU (10 μM) alone and associated with 25 μM resveratrol combination, then applied to ChIP assay to quantify the STAT3 binding. (D) DLD1 was treated with 5-FU and resveratrol alone and in combination and applied to TRAP-PCR-Elisa assay to quantify telomerase activities. Error bars represent standard deviation. Telomerase activities were measured three times independently.

    Journal: Oncotarget

    Article Title: Combination of resveratrol and 5-flurouracil enhanced anti-telomerase activity and apoptosis by inhibiting STAT3 and Akt signaling pathways in human colorectal cancer cells

    doi: 10.18632/oncotarget.25993

    Figure Lengend Snippet: STAT3 binding to promoter region hTERT were inhibited by the resveratrol and 5-FU alone and in combination and corresponding telomerase activities were decreased upon the combination treatments Transcription factor STAT3 binding to hTERT promoter region was tested with chromatin immunoprecipitation assay. (A) HCT116 was treated with 5-FU and resveratrol alone and in combination and applied to ChIP assay. STAT3 binding was quantified by the ChIP band intensity relative to untreated control, which was set as 1.0. (B) Telomerase activities were measured by TRAP PCR reaction conjugated to Elisa assay. (C) DLD1 was treated with 5-FU (10 μM) alone and associated with 25 μM resveratrol combination, then applied to ChIP assay to quantify the STAT3 binding. (D) DLD1 was treated with 5-FU and resveratrol alone and in combination and applied to TRAP-PCR-Elisa assay to quantify telomerase activities. Error bars represent standard deviation. Telomerase activities were measured three times independently.

    Article Snippet: Telomerase activity assays Cancer cells were processed according to the manufacturer's protocol for the TeloTAGGG Telomerase PCR ELISA kit (Roche, Orange, CA.

    Techniques: Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Schematic representation of combination treatment effects with 5-FU and resveratrol in colorectal cancer cells The combined treatments with 5-FU and resveratrol inhibit Akt and STAT3 signaling pathways in colorectal cancer. Akt plays a key role in cell proliferation and survival and STAT3 is key transcription factor for telomerase and other target genes involved in immune response and potential stem-like traits. Our model suggests that combined treatments of 5-FU and resveratrol drive apoptosis by inhibiting Akt and STAT3, concurrently decreasing telomerase activity and downregulating target genes of STAT3 leading to re-sensitization of colorectal cancer to chemotherapy.

    Journal: Oncotarget

    Article Title: Combination of resveratrol and 5-flurouracil enhanced anti-telomerase activity and apoptosis by inhibiting STAT3 and Akt signaling pathways in human colorectal cancer cells

    doi: 10.18632/oncotarget.25993

    Figure Lengend Snippet: Schematic representation of combination treatment effects with 5-FU and resveratrol in colorectal cancer cells The combined treatments with 5-FU and resveratrol inhibit Akt and STAT3 signaling pathways in colorectal cancer. Akt plays a key role in cell proliferation and survival and STAT3 is key transcription factor for telomerase and other target genes involved in immune response and potential stem-like traits. Our model suggests that combined treatments of 5-FU and resveratrol drive apoptosis by inhibiting Akt and STAT3, concurrently decreasing telomerase activity and downregulating target genes of STAT3 leading to re-sensitization of colorectal cancer to chemotherapy.

    Article Snippet: Telomerase activity assays Cancer cells were processed according to the manufacturer's protocol for the TeloTAGGG Telomerase PCR ELISA kit (Roche, Orange, CA.

    Techniques: Activity Assay

    Comparison of HBV replication between wild-type (WT) and mutants SPII in HepG2 cells. The p1.2/PC-based SPII WT and mutant plasmids were transfected into HepG2 cells that were harvested 72 hours after transfection. Total RNA was isolated for detection of HBV total RNA ( A ) and pregenomic RNA (pgRNA) ( B ) using RT-qPCR. HBV DNA from cell supernatant ( C ) and cell lysate ( D ) was measured by qPCR. Data shown as fold change relative to WT. Mann–Whitney U tests were employed. ** p

    Journal: Viruses

    Article Title: Naturally Occurring Mutations within HBV Surface Promoter II Sequences Affect Transcription Activity, HBsAg and HBV DNA Levels in HBeAg-Positive Chronic Hepatitis B Patients

    doi: 10.3390/v11010078

    Figure Lengend Snippet: Comparison of HBV replication between wild-type (WT) and mutants SPII in HepG2 cells. The p1.2/PC-based SPII WT and mutant plasmids were transfected into HepG2 cells that were harvested 72 hours after transfection. Total RNA was isolated for detection of HBV total RNA ( A ) and pregenomic RNA (pgRNA) ( B ) using RT-qPCR. HBV DNA from cell supernatant ( C ) and cell lysate ( D ) was measured by qPCR. Data shown as fold change relative to WT. Mann–Whitney U tests were employed. ** p

    Article Snippet: The detection limit of plasma HBV DNA by the Roche TaqMan48 automatic florescence quantitative polymerase chain reaction (qPCR) kit was 12 IU/mL.

    Techniques: Mutagenesis, Transfection, Isolation, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, MANN-WHITNEY

    Scatter plots presenting levels of HBsAg ( A )/HBV DNA ( B ) for 87 chronic hepatitis B (CHB) C2-subgenotype patients without/with G120A mutation. According to Mann–Whitney U tests, the virological characteristics between the two groups were statistically different ( p = 0.0040 for HBsAg and p = 0.0237 for HBV DNA, respectively).

    Journal: Viruses

    Article Title: Naturally Occurring Mutations within HBV Surface Promoter II Sequences Affect Transcription Activity, HBsAg and HBV DNA Levels in HBeAg-Positive Chronic Hepatitis B Patients

    doi: 10.3390/v11010078

    Figure Lengend Snippet: Scatter plots presenting levels of HBsAg ( A )/HBV DNA ( B ) for 87 chronic hepatitis B (CHB) C2-subgenotype patients without/with G120A mutation. According to Mann–Whitney U tests, the virological characteristics between the two groups were statistically different ( p = 0.0040 for HBsAg and p = 0.0237 for HBV DNA, respectively).

    Article Snippet: The detection limit of plasma HBV DNA by the Roche TaqMan48 automatic florescence quantitative polymerase chain reaction (qPCR) kit was 12 IU/mL.

    Techniques: Mutagenesis, MANN-WHITNEY

    Antibody competition titration assays using MSP1 19 proteins from four Plasmodium species. A combined dilution (1:400 of each serum) containing sera from chimpanzees experimentally infected with either P. malariae (Klimatis), P. ovale (Alpert) or P. vivax (Duff) was incubated with the indicated concentrations of the MSP1 19 competitor protein for 1 h at room temperature. Competitor proteins used were: a P. falciparum MSP1 19 ; b P. malariae MSP1 19 ; c P. ovale MSP1 19 ; d P. vivax MSP1 19 . Multiplex bead assays were performed as described in “ Methods ” and the multiplex response in MFI-bg units are plotted versus the competitor concentration. Multiplex responses are presented as a percentage of the assay results for the PBS control

    Journal: Malaria Journal

    Article Title: Specificity of the IgG antibody response to Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale MSP119 subunit proteins in multiplexed serologic assays

    doi: 10.1186/s12936-018-2566-0

    Figure Lengend Snippet: Antibody competition titration assays using MSP1 19 proteins from four Plasmodium species. A combined dilution (1:400 of each serum) containing sera from chimpanzees experimentally infected with either P. malariae (Klimatis), P. ovale (Alpert) or P. vivax (Duff) was incubated with the indicated concentrations of the MSP1 19 competitor protein for 1 h at room temperature. Competitor proteins used were: a P. falciparum MSP1 19 ; b P. malariae MSP1 19 ; c P. ovale MSP1 19 ; d P. vivax MSP1 19 . Multiplex bead assays were performed as described in “ Methods ” and the multiplex response in MFI-bg units are plotted versus the competitor concentration. Multiplex responses are presented as a percentage of the assay results for the PBS control

    Article Snippet: Comparison of Plasmodium malariae MSP119 sequences from other geographic locations Ten nanograms of DNA from P. malariae strains Greece I, Guyana, and Uganda I were PCR amplified using the forward and reverse long deoxyoligonucleotides described above and the Expand High Fidelity PCR system (Roche Applied Science, Indianapolis, IN, USA).

    Techniques: Titration, Infection, Incubation, Multiplex Assay, Concentration Assay

    Antibody competition titration assays using homologous MSP1 19 proteins. Dilutions (1:400) of P. falciparum Lot 6 defined human serum or of sera from chimpanzees experimentally infected with either P. malariae (Klimatis), P. ovale (Alpert) or P. vivax (Duff) were incubated with the indicated concentrations of the homologous MSP1 19 competitor protein for 1 h at room temperature. Multiplex bead assays were performed as described in “ Methods ”, and the multiplex responses in MFI-bg units are plotted versus the competitor concentration

    Journal: Malaria Journal

    Article Title: Specificity of the IgG antibody response to Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale MSP119 subunit proteins in multiplexed serologic assays

    doi: 10.1186/s12936-018-2566-0

    Figure Lengend Snippet: Antibody competition titration assays using homologous MSP1 19 proteins. Dilutions (1:400) of P. falciparum Lot 6 defined human serum or of sera from chimpanzees experimentally infected with either P. malariae (Klimatis), P. ovale (Alpert) or P. vivax (Duff) were incubated with the indicated concentrations of the homologous MSP1 19 competitor protein for 1 h at room temperature. Multiplex bead assays were performed as described in “ Methods ”, and the multiplex responses in MFI-bg units are plotted versus the competitor concentration

    Article Snippet: Comparison of Plasmodium malariae MSP119 sequences from other geographic locations Ten nanograms of DNA from P. malariae strains Greece I, Guyana, and Uganda I were PCR amplified using the forward and reverse long deoxyoligonucleotides described above and the Expand High Fidelity PCR system (Roche Applied Science, Indianapolis, IN, USA).

    Techniques: Titration, Infection, Incubation, Multiplex Assay, Concentration Assay

    Alignment of predicted Plasmodium spp. MSP1 19 protein sequences using COBALT [ 61 ]. Residues in the P. malariae sequence that differ from the Cameroon sequence of Birkenmeyer et al. [ 38 ] are shaded. Predicted protein sequences resulting from the oligonucleotides used in PCR amplification are underlined. The positions of residues conserved among all the presented MSP1 19 protein sequences are indicated in the consensus with divergent residues indicated by a dot. GenBank accession numbers are MH577181, P. ovale Nigeria I strain; MH577182, P. malariae China I strain; MH577183, P. malariae Greece I strain; MH577184, P. malariae Uganda I strain; and MH577185, P. malariae Guyana strain

    Journal: Malaria Journal

    Article Title: Specificity of the IgG antibody response to Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale MSP119 subunit proteins in multiplexed serologic assays

    doi: 10.1186/s12936-018-2566-0

    Figure Lengend Snippet: Alignment of predicted Plasmodium spp. MSP1 19 protein sequences using COBALT [ 61 ]. Residues in the P. malariae sequence that differ from the Cameroon sequence of Birkenmeyer et al. [ 38 ] are shaded. Predicted protein sequences resulting from the oligonucleotides used in PCR amplification are underlined. The positions of residues conserved among all the presented MSP1 19 protein sequences are indicated in the consensus with divergent residues indicated by a dot. GenBank accession numbers are MH577181, P. ovale Nigeria I strain; MH577182, P. malariae China I strain; MH577183, P. malariae Greece I strain; MH577184, P. malariae Uganda I strain; and MH577185, P. malariae Guyana strain

    Article Snippet: Comparison of Plasmodium malariae MSP119 sequences from other geographic locations Ten nanograms of DNA from P. malariae strains Greece I, Guyana, and Uganda I were PCR amplified using the forward and reverse long deoxyoligonucleotides described above and the Expand High Fidelity PCR system (Roche Applied Science, Indianapolis, IN, USA).

    Techniques: Sequencing, Polymerase Chain Reaction, Amplification

    Colocalization and effects of direct NPFFR2 signalling on NPY neurons. a Representative image of GFP expression in the Arc of a NPY-TRAP mouse brain. Scale bar = 100 µm. b, c Quantification of the expression of Npy and Npffr2 mRNA in the input and immunoprecipitated (IP) RNA isolated from the Arc of NPY-TRAP ( n = 10), Ins-TRAP ( n = 3) and WT-TRAP ( n = 3) mice. One-way ANOVA was used to determine difference between groups. ∗∗ p

    Journal: Nature Communications

    Article Title: Diet-induced adaptive thermogenesis requires neuropeptide FF receptor-2 signalling

    doi: 10.1038/s41467-018-06462-0

    Figure Lengend Snippet: Colocalization and effects of direct NPFFR2 signalling on NPY neurons. a Representative image of GFP expression in the Arc of a NPY-TRAP mouse brain. Scale bar = 100 µm. b, c Quantification of the expression of Npy and Npffr2 mRNA in the input and immunoprecipitated (IP) RNA isolated from the Arc of NPY-TRAP ( n = 10), Ins-TRAP ( n = 3) and WT-TRAP ( n = 3) mice. One-way ANOVA was used to determine difference between groups. ∗∗ p

    Article Snippet: RT-qPCR using primers for Npy , GFP and Npffr2 was carried out in samples prior (input) and after the immunoprecipitation in at least triplicates from 1:5 dilution cDNA from each sample using the LightCycler® (LightCycler® 480 Real-Time PCR system, Roche Applied Science, Germany), SYBR Green I (Molecular Probes) and Platinum Taq DNA Polymerase (Invitrogen).

    Techniques: Expressing, Immunoprecipitation, Isolation, Mouse Assay