Structured Review

Promega promega kit
Emax NGF <t>Promega:</t> NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.
Promega Kit, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/promega kit/product/Promega
Average 99 stars, based on 19 article reviews
Price from $9.99 to $1999.99
promega kit - by Bioz Stars, 2020-03
99/100 stars

Images

1) Product Images from "NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward"

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

Journal: Frontiers in Molecular Neuroscience

doi: 10.3389/fnmol.2016.00063

Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.
Figure Legend Snippet: Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.

Techniques Used:

NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p
Figure Legend Snippet: NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p

Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay

2) Product Images from "Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells"

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells

Journal: International Journal of Biological Sciences

doi:

PP2A shows opposing activity with diffuse alternations of its core enzyme protein level and post-translational modification under MCLR treatment. HEK293 cells were exposed to varying concentrations of MCLR for 24 h before PP2A analysis. (A) PP2A activity was measured with a Promega serine/threonine phosphatase kit assay. Data represent means ±SD (n=3) *, p
Figure Legend Snippet: PP2A shows opposing activity with diffuse alternations of its core enzyme protein level and post-translational modification under MCLR treatment. HEK293 cells were exposed to varying concentrations of MCLR for 24 h before PP2A analysis. (A) PP2A activity was measured with a Promega serine/threonine phosphatase kit assay. Data represent means ±SD (n=3) *, p

Techniques Used: Activity Assay, Modification

3) Product Images from "Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients"

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients

Journal: Cancer Biology & Medicine

doi: 10.20892/j.issn.2095-3941.2016.0091

Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).
Figure Legend Snippet: Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).

Techniques Used: Marker

Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).
Figure Legend Snippet: Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).

Techniques Used: Immunohistochemistry, Staining, Expressing

Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).
Figure Legend Snippet: Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).

Techniques Used: Electrophoresis, Marker, Multiplex Assay, Polymerase Chain Reaction, Fluorescence, Amplification

4) Product Images from "NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward"

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

Journal: Frontiers in Molecular Neuroscience

doi: 10.3389/fnmol.2016.00063

Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.
Figure Legend Snippet: Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.

Techniques Used:

NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p
Figure Legend Snippet: NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p

Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay

5) Product Images from "NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward"

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

Journal: Frontiers in Molecular Neuroscience

doi: 10.3389/fnmol.2016.00063

Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.
Figure Legend Snippet: Emax NGF Promega: NGF Standard curves . NGF standard curves carried out by using rat mAb Promega (triangles) and mAb αD11 (Cattaneo et al., 1988 ) (squares) as primary antibody.

Techniques Used:

NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p
Figure Legend Snippet: NGF and proNGF spiked in the sample buffer (supplied by each supplier of the commercial kits) . The histograms summarize the results of the NGF and proNGF spiked separately and together into the assay buffer of the three different commercial kits for the detection of NGF analyzed. The Promega kit was tested in the two different formats: rat mAb Promega and αD11. Two concentrations of recombinant NGF (10 and 100 pg/ml) and proNGF (50 and 200 pg/ml) were spiked into the assay buffer either alone or together with NGF and measured by using the kits previously described. (A) (Emax Promega with mAb Promega), (B) (Emax Promega with mAb αD11), (C) (Chemikine Chemicon), (D) (NGF Rapid ELISA Biosensis) report the values of concentration, interpolated by the calibration curves. The calculated values of the samples spiked with both NGF and proNGF were compared to the spiked value of only NGF. The t -student test was carried out and the p -values were calculated. Two asterisks on the histograms mean a p

Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay

6) Product Images from "Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients"

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients

Journal: Cancer Biology & Medicine

doi: 10.20892/j.issn.2095-3941.2016.0091

Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).
Figure Legend Snippet: Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).

Techniques Used: Marker

Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).
Figure Legend Snippet: Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).

Techniques Used: Immunohistochemistry, Staining, Expressing

Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).
Figure Legend Snippet: Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).

Techniques Used: Electrophoresis, Marker, Multiplex Assay, Polymerase Chain Reaction, Fluorescence, Amplification

7) Product Images from "Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients"

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients

Journal: Cancer Biology & Medicine

doi: 10.20892/j.issn.2095-3941.2016.0091

Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).
Figure Legend Snippet: Number of tumors that were unstable or stable for each of 6 markers (5 markers in Promega kit plus a single CAT25 marker). For CAT25, 25 patients are grouped in MSS (orange) and 6 in MSI-H (blue).

Techniques Used: Marker

Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).
Figure Legend Snippet: Promega system results and IHC for MSH6-deficient patient. Stability in tumor tissue (A) compared with normal tissue (B). IHC staining demonstrating loss of MSH6 expression in the patient (C) (10 ×).

Techniques Used: Immunohistochemistry, Staining, Expressing

Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).
Figure Legend Snippet: Capillary electrophoresis results from the Promega system. A panel consisting of five mononucleotide marker was used for MSI determination via multiplex PCR. X axis is the size in bases. Y axis is the fluorescence intensity. Green peaks are amplification products from microsatellite loci, including three of five markers shown here (NR21, BAT25, and MONO27). Note the shift in the size (bases) of the amplification products in the tumor specimen compared with normal ones. An example of one shifted locus is demonstrated in the tumor sample (B) compared with normal one (A).

Techniques Used: Electrophoresis, Marker, Multiplex Assay, Polymerase Chain Reaction, Fluorescence, Amplification

8) Product Images from "Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing"

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing

Journal: PLoS ONE

doi: 10.1371/journal.pone.0135058

DNA degradation. DNA recovery after bisulfite treatment was determined with the Qubit fluorometer. DNA recovery with the Epigentek kit was 33.2% ± 3.4%, with the Promega kit was 52% ± 3%, with the Diagenode kit was 55% ± 2.6% and with the Qiagen kit was 50.2% ± 2%. The BisulFlash DNA Modification kit was shown to have a significantly decreased average yield with respect to all other kits (FWER
Figure Legend Snippet: DNA degradation. DNA recovery after bisulfite treatment was determined with the Qubit fluorometer. DNA recovery with the Epigentek kit was 33.2% ± 3.4%, with the Promega kit was 52% ± 3%, with the Diagenode kit was 55% ± 2.6% and with the Qiagen kit was 50.2% ± 2%. The BisulFlash DNA Modification kit was shown to have a significantly decreased average yield with respect to all other kits (FWER

Techniques Used: Modification

Related Articles

Clone Assay:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: PCR products were cloned into multiple cloning sites of PmirGLO (Promega Corp., Madison, WI, USA) luciferase gene downstream. .. Dual luciferase activity was detected based on the Promega kit.

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: On the other hand, the algorithms of ESME permit the quantitative analysis directly from four dye trace without the need for cloning, through alignment to a genomic reference sequence, trace off-set correction and normalization of signal intensities. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Luciferase:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: .. Dual luciferase activity was detected based on the Promega kit. .. Western-blotting The transfected cells were collected, washed in phosphate buffered saline (PBS) twice, lysed in 100 μl cell lysate (4°C, 30 min) and centrifuged (12000 g, 10 min) to obtain the supernatant, and the protein concentration was determined by Bradford method.

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: .. Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA). ..

Phosphatase Assay:

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit. .. To confirm this result, we tested PP2A activity at 2μM and 10μM MCLR treatment, and control, by an immunoprecipitation phosphatase assay method.

Synthesized:

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: Single-stranded 43-mer anti-sense oligonucleotide (10 pmol) 5′-CTCTCGCCACTGCGCTGCGCTTCTCCTCTCAGGGTGCCGTCGC-3′ (Tm = 75.3°C) corresponding to exon 1 in the P2Y2 R gene sequence was designed and chemically synthesized, and end-labeled using polynucleotide kinase from Promega, and [ γ -32 P]EasyTides ATP was from PerkinElmer. .. Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control.

Real-time Polymerase Chain Reaction:

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: Paragraph title: RNA extraction and real-time PCR ... Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI).

Sandwich ELISA:

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: In order to verify the obtained results, two other commercial kits (ChemiKine Nerve Growth Factor, Sandwich ELISA, from Millipore, and Mouse NGF Rapid ELISA Kit from Biosensis) were tested. .. The experiments were repeated three times for Promega kit, with both the antibodies, two times for Biosensis and Millipore kits.

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: .. In order to verify whether this interference was due to the specific layout of the Promega kit, the NGF/proNGF spiking experiments were carried out also with two other commercial kits: ChemiKine NGF Sandwich ELISA (Millipore), and Mouse NGF Rapid ELISA (Biosensis). .. With both assays, proNGF is poorly detected if tested alone, while NGF alone is largely overestimated, especially by the Biosensis ELISA (Table ).

Incubation:

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: The MCLR concentration and incubation time were decided according to previous studies performed by our, and other, groups - . .. After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit.

Amplification:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: Full-length 3′-untranslatedregion (3′-UTR) of the FAK gene was amplified, cloned and sequenced. .. Dual luciferase activity was detected based on the Promega kit.

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: Therefore, these two kits were further used for bisulfite amplicon sequencing on an Illumina MiSeq platform. shows the observed methylation levels in correlation with the expected methylation levels. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Activity Assay:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: .. Dual luciferase activity was detected based on the Promega kit. .. Western-blotting The transfected cells were collected, washed in phosphate buffered saline (PBS) twice, lysed in 100 μl cell lysate (4°C, 30 min) and centrifuged (12000 g, 10 min) to obtain the supernatant, and the protein concentration was determined by Bradford method.

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: .. After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit. .. To confirm this result, we tested PP2A activity at 2μM and 10μM MCLR treatment, and control, by an immunoprecipitation phosphatase assay method.

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: .. Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA). ..

Expressing:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: The binding sites of miR-7 and target genes were subjected to site-specific mutagenesis, and pRL-TK vector expressing Renilla luciferase (TaKaRa) was used as internal reference to adjust differences in cell numbers and transfection efficiency. .. Dual luciferase activity was detected based on the Promega kit.

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA). .. LucSV40 (Luc-4) was used as a reference for maximal luciferase activity, as it contains the SV40 promoter and enhancer, while Luc-1 lacking regulatory sequences was used as a negative control to determine the basal non-specific luciferase expression, which was found to be negligible.

Bradford Assay:

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: The concentration of the total protein in the sample, measured by Bradford assay (Sigma-Aldrich), was 27 mg/ml. proNGF at the same concentrations assayed in assay buffer, were spiked into TgProNGF#72 homogenate (1:25, 1:50, 1:100, and 1:200), and measured. .. The experiments were repeated three times for Promega kit, with both the antibodies, two times for Biosensis and Millipore kits.

Modification:

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: Paragraph title: PP2A activity undergoes opposite change with alternations of the protein level and modification of its subunits ... After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit.

Transfection:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: The binding sites of miR-7 and target genes were subjected to site-specific mutagenesis, and pRL-TK vector expressing Renilla luciferase (TaKaRa) was used as internal reference to adjust differences in cell numbers and transfection efficiency. .. Dual luciferase activity was detected based on the Promega kit.

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: Paragraph title: In vitro transfection and luciferase assay ... Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA).

Positive Control:

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: .. Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control. .. The negative control included diethylpyrocarbonate-treated water instead of mRNA in the reaction.

Concentration Assay:

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: The concentration of the total protein in the sample, measured by Bradford assay (Sigma-Aldrich), was 27 mg/ml. proNGF at the same concentrations assayed in assay buffer, were spiked into TgProNGF#72 homogenate (1:25, 1:50, 1:100, and 1:200), and measured. .. The experiments were repeated three times for Promega kit, with both the antibodies, two times for Biosensis and Millipore kits.

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: A cell density of 6 × 103 cells/mL for fixation and magnetic bead-based DNA extraction using the Promega kit, with any fixation time, resulted in the highest quality and quantity of DNA; the DNA was also of suitable quality for light microscopic LBC observation. .. A higher density of 1 × 105 cells/mL resulted in a reduced concentration of amplifiable DNA, indicating that the DNA extraction kits displayed certain use limitations in our experimental conditions.

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: .. After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit. .. To confirm this result, we tested PP2A activity at 2μM and 10μM MCLR treatment, and control, by an immunoprecipitation phosphatase assay method.

MSI Assay:

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: In MSI-L, all CRCs were relatively unstable if enough markers were used in the MSI assay. .. Therefore, detecting MSI-L by more markers, such as Promega kit but not in single marker test, e.g., CAT25 marker that we tested here, is possible.

Biomarker Assay:

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: To find the best marker as a biomarker tool for diagnosis, many investigations have been conducted on MSI markers because MSI is an important marker to screen for HNPCC as well as a prognostic and predictive marker for sporadic colorectal cancer - . .. Promega kit offered a highly sensitive and specific method in the detection of MSI.

Cell Culture:

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: Discussion In the present study, we clearly demonstrated that DNA obtained from cultured cells and clinical LBC specimens fixed with CytoRich Red solution were suitable for cancer genome analysis using NGS, even after the samples were stored for up to 1 year. .. A cell density of 6 × 103 cells/mL for fixation and magnetic bead-based DNA extraction using the Promega kit, with any fixation time, resulted in the highest quality and quantity of DNA; the DNA was also of suitable quality for light microscopic LBC observation.

Generated:

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: The data generated from the Premium Bisulfite kit and the MethylEdge Bisulfite Conversion System showed higher correlation coefficients suggesting more accurate methylation quantitation. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Sequencing:

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: Single-stranded 43-mer anti-sense oligonucleotide (10 pmol) 5′-CTCTCGCCACTGCGCTGCGCTTCTCCTCTCAGGGTGCCGTCGC-3′ (Tm = 75.3°C) corresponding to exon 1 in the P2Y2 R gene sequence was designed and chemically synthesized, and end-labeled using polynucleotide kinase from Promega, and [ γ -32 P]EasyTides ATP was from PerkinElmer. .. Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control.

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI). .. Cytokine primer and probe sequences were developed with Primer Express Software from PE Biosystems (Foster City, CA), and the influenza A matrix (M1) protein primers and probe were based on a previously published sequence ( ).

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: Three categories of MSI status are observed by using Promega kit, MSI-H, MSI-L, and MSS showing more or less than 30% instabilities of the markers. .. Forward primer sequence is 5′-CCTAG AAACCTTTATCCCTGCTT-3′ and reverse primer sequence is 5′-GAGCTTGCAGTGAGCTGAGA-3′.

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: Therefore, these two kits were further used for bisulfite amplicon sequencing on an Illumina MiSeq platform. shows the observed methylation levels in correlation with the expected methylation levels. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Binding Assay:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: The binding sites of miR-7 and target genes were subjected to site-specific mutagenesis, and pRL-TK vector expressing Renilla luciferase (TaKaRa) was used as internal reference to adjust differences in cell numbers and transfection efficiency. .. Dual luciferase activity was detected based on the Promega kit.

Reporter Gene Assay:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: Paragraph title: Dual luciferase reporter gene assay ... Dual luciferase activity was detected based on the Promega kit.

Molecular Weight:

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: Transient transfections were carried out using the JetPEI cationic polymer transfection reagent (mean molecular weight of 22 kDa; Polyplus, Illkirsh, France). .. Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA).

DNA Extraction:

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: .. A cell density of 6 × 103 cells/mL for fixation and magnetic bead-based DNA extraction using the Promega kit, with any fixation time, resulted in the highest quality and quantity of DNA; the DNA was also of suitable quality for light microscopic LBC observation. .. A higher density of 1 × 105 cells/mL resulted in a reduced concentration of amplifiable DNA, indicating that the DNA extraction kits displayed certain use limitations in our experimental conditions.

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: .. The residual LBC breast cancer samples collected from Hokuto Hospital that were initially obtained by FNA and fixed with CytoRich Red solution were subjected to DNA extraction using the Promega kit and processed for NGS using the QIAseq Human Breast Cancer Panel. ..

Methylation:

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: Therefore, detecting MSI-L by more markers, such as Promega kit but not in single marker test, e.g., CAT25 marker that we tested here, is possible. .. High frequency of K-RAS mutations in MSI-L cancers and methylation of methylguanine transferase in MSI-L tumors have been reported in contrast to MSI-H where more mutations occur in MMR pathway; however, this finding is not clear in others - .

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: Therefore, these two kits were further used for bisulfite amplicon sequencing on an Illumina MiSeq platform. shows the observed methylation levels in correlation with the expected methylation levels. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Mutagenesis:

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: The binding sites of miR-7 and target genes were subjected to site-specific mutagenesis, and pRL-TK vector expressing Renilla luciferase (TaKaRa) was used as internal reference to adjust differences in cell numbers and transfection efficiency. .. Dual luciferase activity was detected based on the Promega kit.

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: Comprehensive genomic DNA analysis by NGS using cells fixed with CytoRich Red solution DNA extracted from MCF-7 cells were subjected to NGS using the QIAseq Human Breast Cancer Panel with DNA extracted using the Promega kit, in which NGS results showed the same gene mutation variants as previously reported [ ] ( ). .. The residual LBC breast cancer samples collected from Hokuto Hospital that were initially obtained by FNA and fixed with CytoRich Red solution were subjected to DNA extraction using the Promega kit and processed for NGS using the QIAseq Human Breast Cancer Panel.

Isolation:

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: Cortex brain areas (CTX) were isolated from TgProNGF#72mice, 12 month old. .. The experiments were repeated three times for Promega kit, with both the antibodies, two times for Biosensis and Millipore kits.

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: Labeled primers (1 pmol) were hybridized for 20 min with 5 μ g of polyadenylate+ RNA isolated from HCAECs, and avian myeloblastosis virus reverse transcriptase was added for 30 min at 41°C to yield the corresponding cDNA. .. Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control.

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: RNA was isolated from the apical lobe of the lung in TRIzol, according to manufacturer’s instructions (Life Technologies, Carlsbad, CA). .. Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI).

Negative Control:

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control. .. The negative control included diethylpyrocarbonate-treated water instead of mRNA in the reaction.

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA). .. LucSV40 (Luc-4) was used as a reference for maximal luciferase activity, as it contains the SV40 promoter and enhancer, while Luc-1 lacking regulatory sequences was used as a negative control to determine the basal non-specific luciferase expression, which was found to be negligible.

Labeling:

Article Title: P2Y2 Receptor Transcription Is Increased by NF-κB and Stimulates Cyclooxygenase-2 Expression and PGE2 Released by Intestinal Epithelial Cells
Article Snippet: Labeled primers (1 pmol) were hybridized for 20 min with 5 μ g of polyadenylate+ RNA isolated from HCAECs, and avian myeloblastosis virus reverse transcriptase was added for 30 min at 41°C to yield the corresponding cDNA. .. Kanamycin-positive mRNA (1.2 kb) from the the Promega kit was used with a corresponding primer as a positive control.

Article Title: Serotonin 3A Receptor Subtype as an Early and Protracted Marker of Cortical Interneuron Subpopulations
Article Snippet: .. The transcription was carried out using the Promega kit, and probes were labeled with 35 S-UTP ( > 1000 Ci/mmol; Amersham). ..

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI). .. Cytokine and 18s probes were labeled at the 5′ end with the reporter dye FAM and at the 3′ end with the quencher dye TAMRA.

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: Three categories of MSI status are observed by using Promega kit, MSI-H, MSI-L, and MSS showing more or less than 30% instabilities of the markers. .. PCR primers were labeled at the 5′ end with Cy5 for the subsequent analysis using ALFexpress DNA sequencer (Amersham Pharmasia Biotech).

Purification:

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: A cell density of 6 × 103 cells/mL for fixation and magnetic bead-based DNA extraction using the Promega kit, with any fixation time, resulted in the highest quality and quantity of DNA; the DNA was also of suitable quality for light microscopic LBC observation. .. However, the DNA quality was high enough for NGS when DNA was extracted using the other magnetic resin-based DNA extraction kit (Agencourt FormaPure DNA kit), although this kit showed lower extraction efficiency than the Promega purification kit.

Polymerase Chain Reaction:

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: Paragraph title: MSI status by PCR-based methods ... Three categories of MSI status are observed by using Promega kit, MSI-H, MSI-L, and MSS showing more or less than 30% instabilities of the markers.

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: PCR products were cloned into multiple cloning sites of PmirGLO (Promega Corp., Madison, WI, USA) luciferase gene downstream. .. Dual luciferase activity was detected based on the Promega kit.

In Situ Hybridization:

Article Title: Serotonin 3A Receptor Subtype as an Early and Protracted Marker of Cortical Interneuron Subpopulations
Article Snippet: Paragraph title: Radioactive In Situ Hybridization ... The transcription was carried out using the Promega kit, and probes were labeled with 35 S-UTP ( > 1000 Ci/mmol; Amersham).

Plasmid Preparation:

Article Title: Serotonin 3A Receptor Subtype as an Early and Protracted Marker of Cortical Interneuron Subpopulations
Article Snippet: The plasmid was linearized with Bam H1 for antisense RNA synthesis by T7 polymerase and with Eco RI for sense RNA synthesis by T3 polymerase. .. The transcription was carried out using the Promega kit, and probes were labeled with 35 S-UTP ( > 1000 Ci/mmol; Amersham).

Article Title: MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway
Article Snippet: MiR-7 mimics and miR-7 mimic NC were respectively co-transfected with luciferase reporter vector into NSCLC cells. .. Dual luciferase activity was detected based on the Promega kit.

Software:

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI). .. Cytokine primer and probe sequences were developed with Primer Express Software from PE Biosystems (Foster City, CA), and the influenza A matrix (M1) protein primers and probe were based on a previously published sequence ( ).

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: Three categories of MSI status are observed by using Promega kit, MSI-H, MSI-L, and MSS showing more or less than 30% instabilities of the markers. .. Primer-BLAST software was used to design primers for CAT25 locus.

Enzyme-linked Immunosorbent Assay:

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: Paragraph title: ProNGF interferes with the determination of NGF by using commercial ELISA ... The experiments were repeated three times for Promega kit, with both the antibodies, two times for Biosensis and Millipore kits.

Article Title: NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward
Article Snippet: .. In order to verify whether this interference was due to the specific layout of the Promega kit, the NGF/proNGF spiking experiments were carried out also with two other commercial kits: ChemiKine NGF Sandwich ELISA (Millipore), and Mouse NGF Rapid ELISA (Biosensis). .. With both assays, proNGF is poorly detected if tested alone, while NGF alone is largely overestimated, especially by the Biosensis ELISA (Table ).

RNA Extraction:

Article Title: Influenza Virus-Specific Immunological Memory Is Enhanced by Repeated Social Defeat
Article Snippet: Paragraph title: RNA extraction and real-time PCR ... Reverse transcription and cDNA synthesis were carried out using a Promega kit (Madison, WI).

In Vitro:

Article Title: Development of targeted therapy for ovarian cancer mediated by a plasmid expressing diphtheria toxin under the control of H19 regulatory sequences
Article Snippet: Paragraph title: In vitro transfection and luciferase assay ... Luciferase activity was measured using the Promega kit 'Luciferase Assay System' (E-1500; Promega, Madison, USA).

Next-Generation Sequencing:

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: The DNA quality of DNA extracted using four different DNA extraction methods was comparable, and all the samples were suitable for the construction of NGS libraries and for cancer genome analysis. .. A cell density of 6 × 103 cells/mL for fixation and magnetic bead-based DNA extraction using the Promega kit, with any fixation time, resulted in the highest quality and quantity of DNA; the DNA was also of suitable quality for light microscopic LBC observation.

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: .. The residual LBC breast cancer samples collected from Hokuto Hospital that were initially obtained by FNA and fixed with CytoRich Red solution were subjected to DNA extraction using the Promega kit and processed for NGS using the QIAseq Human Breast Cancer Panel. ..

Quantitation Assay:

Article Title: Bisulfite Conversion of DNA: Performance Comparison of Different Kits and Methylation Quantitation of Epigenetic Biomarkers that Have the Potential to Be Used in Non-Invasive Prenatal Testing
Article Snippet: The data generated from the Premium Bisulfite kit and the MethylEdge Bisulfite Conversion System showed higher correlation coefficients suggesting more accurate methylation quantitation. .. The λ-DNA control was able to fit linear lines with both the Promega and Diagenode kit with slightly higher corelation coefficient when using the Promega kit.

Immunoprecipitation:

Article Title: Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by ?4 Protein in HEK293 Cells
Article Snippet: After peaking at a 2 μM concentration at a rise of about 30%, PP2A activity decreased in a dose-dependent manner to about 20% for a 10 μM concentration (Fig. A), measured by a Promega kit. .. To confirm this result, we tested PP2A activity at 2μM and 10μM MCLR treatment, and control, by an immunoprecipitation phosphatase assay method.

Marker:

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: .. Therefore, detecting MSI-L by more markers, such as Promega kit but not in single marker test, e.g., CAT25 marker that we tested here, is possible. ..

Article Title: Simplified microsatellite instability detection protocol provides equivalent sensitivity to robust detection strategies in Lynch syndrome patients
Article Snippet: To find the best marker as a biomarker tool for diagnosis, many investigations have been conducted on MSI markers because MSI is an important marker to screen for HNPCC as well as a prognostic and predictive marker for sporadic colorectal cancer - . .. Promega kit offered a highly sensitive and specific method in the detection of MSI.

Variant Assay:

Article Title: Comprehensive validation of liquid-based cytology specimens for next-generation sequencing in cancer genome analysis
Article Snippet: The NGS results, including mean read depth and variant allele fraction, were comparable, and variant calls were exactly same across the three different DNA extraction methods in each cell line ( ). .. The residual LBC breast cancer samples collected from Hokuto Hospital that were initially obtained by FNA and fixed with CytoRich Red solution were subjected to DNA extraction using the Promega kit and processed for NGS using the QIAseq Human Breast Cancer Panel.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • pna  (Promega)
    93
    Promega pna
    Evaluation of maximum sensitivity of <t>PNA</t> method using oligo targets containing minimal base pair alteration. a and b PNA analysis was performed using oligo samples containing different portions of <t>MSI</t> variants with two-base deletion or one-base insertion. PNA method clearly detected two-base deletion and one-base insertion in all five MSI markers in samples containing 5% or more MSI-H variant
    Pna, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pna/product/Promega
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    pna - by Bioz Stars, 2020-03
    93/100 stars
      Buy from Supplier

    79
    Promega in vitro ced 3 zymogen activation assay
    CSP-3 specifically inhibits the autoactivation of the <t>CED-3</t> zymogen ( a ) CSP-3 inhibits autoactivation of the CED-3 zymogen. GST (93 nM, lanes 1–3), GST–CSP-3 (12 nM, lanes 4–6) or GST–CSP-3(F57D) (12 nM, lanes 7–9) was incubated with 35 S-methionine labeled CED-3 zymogen as described in Methods . At different time points, an aliquot was taken out and SDS sampling buffer was added to stop the reaction. The samples were resolved by 15% SDS-PAGE and subjected to autoradiography. ( b ) CSP-3 delays but does not block CED-4–mediated activation of CED-3. GST (93 nM) or GST–CSP-3 (12 nM) was incubated with 35 S-methionine labeled CED-3 zymogen in the absence or presence of oligomeric CED-4 (40 nM) (added 20 min later). At different time points, an aliquot was taken out and SDS sampling buffer was added. One half of the aliquot was resolved by 15% SDS-PAGE and subjected to autoradiography (above). One-fourth of the aliquot was used for the immunoblotting analysis using an anti-GST antibody (below). ( c ) Oligomeric CED-4 overrides CSP-3 inhibition to induce CED-3 activation. Preformed GST-CSP-3–CED-3-Flag complexes (~70 nM of CED-3–Flag) were incubated either with buffer or with increasing concentrations of oligomeric or monomeric CED-4 at 30 °C for 30 min (Methods), before being resolved on 15% SDS-PAGE and detected with an anti-Flag antibody. ( d ) CSP-3 does not inhibit the activity of the active CED-3 protease in vitro . CED-3–Flag was coexpressed with GST, GST–CSP-3 or GST–CSP-3(F57D) for 3 h in bacteria. The bacterial lysate containing similar levels of active CED-3 (acCED-3) and GST fusion proteins ( Supplementary Fig. 2b ) was incubated with 35 S-methionine–labeled CED-9 for 2 h at 30 °C. In one reaction, the caspase inhibitor iodoacetic acid (5 mM) was included. The reactions were resolved by 15% SDS-PAGE and detected by autoradiography. ( e ) A CSP-3 working model in C. elegans . CSP-3 associates with the CED-3 zymogen in all cells. In cells that normally live, CSP-3 prevents the CED-3 zymogen from dimerization and inadvertent autoactivation. In cells that are programmed to die, CED-4 oligomer overrides CSP-3 inhibition to induce the activation of the CED-3 zymogen via the proximity-induced dimerization model 6 .
    In Vitro Ced 3 Zymogen Activation Assay, supplied by Promega, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/in vitro ced 3 zymogen activation assay/product/Promega
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    in vitro ced 3 zymogen activation assay - by Bioz Stars, 2020-03
    79/100 stars
      Buy from Supplier

    98
    Promega caspase 3 activity assay kit
    Role of autophagy induction on the inhibition of apoptosis by globular adiponectin in HepG2 cells. (A) Cells were transfected with LC3B siRNA or scrambled control siRNA followed by incubation with gAcrp (1 μg/ml) for 48 h. Cell viability was assessed as described previously. (B) Cells were transfected with LC3B siRNA or scrambled siRNA followed by gAcrp (1 μg/ml) incubation for 48 h. <t>Caspase-3</t> activity was determined as described previously. (C) Cells were pretreated with indicated concentration of rapamycin for 2 h followed by treatment with gAcrp (1 μg/ml) for 48 h. Caspase-3 activity was determined as described previously. Values represent fold increase compared to control and expressed as mean ± SEM (A and C; n=3: B; n=4). * p
    Caspase 3 Activity Assay Kit, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/caspase 3 activity assay kit/product/Promega
    Average 98 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    caspase 3 activity assay kit - by Bioz Stars, 2020-03
    98/100 stars
      Buy from Supplier

    98
    Promega nadp nadph glo assay kit
    PHGDH-derived serine supports the transsulfuration and folate cycles. (a) Serine metabolism via the transsulfuration and folate cycles. 13 C-labelled carbons (●) unlabelled carbons ( ○ ). Gly: Glycine, Ser: Serine, CTH: Cystathionine; HCY: Homocysteine, GSH: Glutathione, γGC: γ-glutamyl cysteine, Glu: Glutamate. (b–e) A549 cells expressing scramble (SCR), NRF2 or PHGDH shRNAs were grown in the presence of U- 13 C-glucose for 24 hours and metabolites were extracted and analysed by LC/MS. (b) Analysis of 13 C-labelling on the glycine component of glutathione. (c) Analysis of 13 C-labeling on the purine metabolite adenosine monophosphate (AMP). (d,e) Analysis of glutathione (d) and AMP (e) labelling in serine high cell lines. Cells were labelled with 13 C-glucose for 24 or 48 hours as indicated. (b,d) PHGDH-derived serine is incorporated into glutathione through the generation of glycine and cysteine. M+0 denotes no carbons labelled, M+2 denotes labelling on the glycine moiety, M+3 denotes labeling on the cysteine moiety, and M+5 denotes labeling on both glycine and cysteine. M+5 labeling was not observed. (c,e) M+5 labelling occurs following ribose-5-phosphate labelling via the pentose phosphate pathway. M+7 labelling is the result of ribose labelling plus either glycine or formyl-THF labelling in the purine ring. M+9 labelling is the result of ribose labelling plus either glycine and formyl-THF labelling in the purine ring. M+0 has no labelled carbons. (f–g) LC/MS analysis of total metabolite levels in the nucleotide (f) and transsulfuration (g) pathways. (h) <t>NADPH/NADP+</t> ratios 4 days after PHGDH knockdown. Results are the average of 3 biological replicates.
    Nadp Nadph Glo Assay Kit, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nadp nadph glo assay kit/product/Promega
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nadp nadph glo assay kit - by Bioz Stars, 2020-03
    98/100 stars
      Buy from Supplier

    Image Search Results


    Evaluation of maximum sensitivity of PNA method using oligo targets containing minimal base pair alteration. a and b PNA analysis was performed using oligo samples containing different portions of MSI variants with two-base deletion or one-base insertion. PNA method clearly detected two-base deletion and one-base insertion in all five MSI markers in samples containing 5% or more MSI-H variant

    Journal: BMC Cancer

    Article Title: Microsatellite instability test using peptide nucleic acid probe-mediated melting point analysis: a comparison study

    doi: 10.1186/s12885-018-5127-6

    Figure Lengend Snippet: Evaluation of maximum sensitivity of PNA method using oligo targets containing minimal base pair alteration. a and b PNA analysis was performed using oligo samples containing different portions of MSI variants with two-base deletion or one-base insertion. PNA method clearly detected two-base deletion and one-base insertion in all five MSI markers in samples containing 5% or more MSI-H variant

    Article Snippet: Cell lines and patient tissue samples For testing the sensitivity of PNA (U-TOP™ MSI Detection Kit) and MNR (Promega MSI Analysis System Kit), HeLa (microsatellite stable, MSS) and SNU-638 (MSI-H) cell lines were used.

    Techniques: Variant Assay

    Analysis of MSI status in 166 CRCs and matched normal samples using PNA (left panel) and MNR method (right panel). Representative MSI analysis results of CRC samples determined as MSI-H or MSS are shown

    Journal: BMC Cancer

    Article Title: Microsatellite instability test using peptide nucleic acid probe-mediated melting point analysis: a comparison study

    doi: 10.1186/s12885-018-5127-6

    Figure Lengend Snippet: Analysis of MSI status in 166 CRCs and matched normal samples using PNA (left panel) and MNR method (right panel). Representative MSI analysis results of CRC samples determined as MSI-H or MSS are shown

    Article Snippet: Cell lines and patient tissue samples For testing the sensitivity of PNA (U-TOP™ MSI Detection Kit) and MNR (Promega MSI Analysis System Kit), HeLa (microsatellite stable, MSS) and SNU-638 (MSI-H) cell lines were used.

    Techniques:

    Maximum sensitivity evaluation of PNA and MNR method. The maximum sensitivity of PNA and MNR methods was evaluated using mixed samples of genomic DNA samples obtained from HeLa (MSS) and SNU-638 (MSI-H) cells. a PNA method was capable of detecting alteration in all five MSI marker genes in samples containing down to 5% MSI-H variant. b MNR method required at least 20% MSI-H variant in samples to detect alteration in all five MSI marker genes

    Journal: BMC Cancer

    Article Title: Microsatellite instability test using peptide nucleic acid probe-mediated melting point analysis: a comparison study

    doi: 10.1186/s12885-018-5127-6

    Figure Lengend Snippet: Maximum sensitivity evaluation of PNA and MNR method. The maximum sensitivity of PNA and MNR methods was evaluated using mixed samples of genomic DNA samples obtained from HeLa (MSS) and SNU-638 (MSI-H) cells. a PNA method was capable of detecting alteration in all five MSI marker genes in samples containing down to 5% MSI-H variant. b MNR method required at least 20% MSI-H variant in samples to detect alteration in all five MSI marker genes

    Article Snippet: Cell lines and patient tissue samples For testing the sensitivity of PNA (U-TOP™ MSI Detection Kit) and MNR (Promega MSI Analysis System Kit), HeLa (microsatellite stable, MSS) and SNU-638 (MSI-H) cell lines were used.

    Techniques: Marker, Variant Assay

    CSP-3 specifically inhibits the autoactivation of the CED-3 zymogen ( a ) CSP-3 inhibits autoactivation of the CED-3 zymogen. GST (93 nM, lanes 1–3), GST–CSP-3 (12 nM, lanes 4–6) or GST–CSP-3(F57D) (12 nM, lanes 7–9) was incubated with 35 S-methionine labeled CED-3 zymogen as described in Methods . At different time points, an aliquot was taken out and SDS sampling buffer was added to stop the reaction. The samples were resolved by 15% SDS-PAGE and subjected to autoradiography. ( b ) CSP-3 delays but does not block CED-4–mediated activation of CED-3. GST (93 nM) or GST–CSP-3 (12 nM) was incubated with 35 S-methionine labeled CED-3 zymogen in the absence or presence of oligomeric CED-4 (40 nM) (added 20 min later). At different time points, an aliquot was taken out and SDS sampling buffer was added. One half of the aliquot was resolved by 15% SDS-PAGE and subjected to autoradiography (above). One-fourth of the aliquot was used for the immunoblotting analysis using an anti-GST antibody (below). ( c ) Oligomeric CED-4 overrides CSP-3 inhibition to induce CED-3 activation. Preformed GST-CSP-3–CED-3-Flag complexes (~70 nM of CED-3–Flag) were incubated either with buffer or with increasing concentrations of oligomeric or monomeric CED-4 at 30 °C for 30 min (Methods), before being resolved on 15% SDS-PAGE and detected with an anti-Flag antibody. ( d ) CSP-3 does not inhibit the activity of the active CED-3 protease in vitro . CED-3–Flag was coexpressed with GST, GST–CSP-3 or GST–CSP-3(F57D) for 3 h in bacteria. The bacterial lysate containing similar levels of active CED-3 (acCED-3) and GST fusion proteins ( Supplementary Fig. 2b ) was incubated with 35 S-methionine–labeled CED-9 for 2 h at 30 °C. In one reaction, the caspase inhibitor iodoacetic acid (5 mM) was included. The reactions were resolved by 15% SDS-PAGE and detected by autoradiography. ( e ) A CSP-3 working model in C. elegans . CSP-3 associates with the CED-3 zymogen in all cells. In cells that normally live, CSP-3 prevents the CED-3 zymogen from dimerization and inadvertent autoactivation. In cells that are programmed to die, CED-4 oligomer overrides CSP-3 inhibition to induce the activation of the CED-3 zymogen via the proximity-induced dimerization model 6 .

    Journal: Nature structural & molecular biology

    Article Title: Inhibition of CED-3 zymogen activation and apoptosis in Caenorhabditis elegans by caspase homolog CSP-3

    doi: 10.1038/nsmb.1488

    Figure Lengend Snippet: CSP-3 specifically inhibits the autoactivation of the CED-3 zymogen ( a ) CSP-3 inhibits autoactivation of the CED-3 zymogen. GST (93 nM, lanes 1–3), GST–CSP-3 (12 nM, lanes 4–6) or GST–CSP-3(F57D) (12 nM, lanes 7–9) was incubated with 35 S-methionine labeled CED-3 zymogen as described in Methods . At different time points, an aliquot was taken out and SDS sampling buffer was added to stop the reaction. The samples were resolved by 15% SDS-PAGE and subjected to autoradiography. ( b ) CSP-3 delays but does not block CED-4–mediated activation of CED-3. GST (93 nM) or GST–CSP-3 (12 nM) was incubated with 35 S-methionine labeled CED-3 zymogen in the absence or presence of oligomeric CED-4 (40 nM) (added 20 min later). At different time points, an aliquot was taken out and SDS sampling buffer was added. One half of the aliquot was resolved by 15% SDS-PAGE and subjected to autoradiography (above). One-fourth of the aliquot was used for the immunoblotting analysis using an anti-GST antibody (below). ( c ) Oligomeric CED-4 overrides CSP-3 inhibition to induce CED-3 activation. Preformed GST-CSP-3–CED-3-Flag complexes (~70 nM of CED-3–Flag) were incubated either with buffer or with increasing concentrations of oligomeric or monomeric CED-4 at 30 °C for 30 min (Methods), before being resolved on 15% SDS-PAGE and detected with an anti-Flag antibody. ( d ) CSP-3 does not inhibit the activity of the active CED-3 protease in vitro . CED-3–Flag was coexpressed with GST, GST–CSP-3 or GST–CSP-3(F57D) for 3 h in bacteria. The bacterial lysate containing similar levels of active CED-3 (acCED-3) and GST fusion proteins ( Supplementary Fig. 2b ) was incubated with 35 S-methionine–labeled CED-9 for 2 h at 30 °C. In one reaction, the caspase inhibitor iodoacetic acid (5 mM) was included. The reactions were resolved by 15% SDS-PAGE and detected by autoradiography. ( e ) A CSP-3 working model in C. elegans . CSP-3 associates with the CED-3 zymogen in all cells. In cells that normally live, CSP-3 prevents the CED-3 zymogen from dimerization and inadvertent autoactivation. In cells that are programmed to die, CED-4 oligomer overrides CSP-3 inhibition to induce the activation of the CED-3 zymogen via the proximity-induced dimerization model 6 .

    Article Snippet: In vitro CED-3 zymogen activation assay The CED-3 zymogen was first synthesized and labeled with 35 S-methionine in the TNT Transcription/Translation coupled system (Promega) at 30 °C as described previously , in the presence of an equal amount of GST–CSP-3, GST–CSP-3(F57D) or GST.

    Techniques: Incubation, Labeling, Sampling, SDS Page, Autoradiography, Blocking Assay, Activation Assay, Inhibition, Activity Assay, In Vitro

    CSP-3 is a cytoplasmic protein with sequence homology to the small subunit of CED-3 ( a ) Sequence alignment of CSP-3 and the small subunits of CED-3 and human caspase-3. Residues that are identical are shaded in yellow and residues that are similar are labeled with pink. Phe57 of CSP-3 is indicated with a blue arrowhead. ( b ) The csp-3 gene structure and deletion mutations. Exons are depicted as boxes and introns as lines. The translated regions of csp-3 are highlighted in blue. Two orange boxes indicate the regions of csp-3 removed by the two csp-3 deletions, respectively. Also shown are a csp-3 translational gfp fusion and a 5.73-kb genomic rescuing fragment ( csp-3 (+)). ( c ) Both csp-3 deletions abolish expression of csp-3 in C. elegans . Immunoblotting for CSP-3 in total worm lysates from N2 , csp-3 ( tm2260 ) and csp-3 ( tm2486 ) animals is shown. The CstF-64 protein was used as a loading control. ( d ) csp-3 is ubiquitously expressed in C. elegans embryos. Differential interference contrast (DIC) and GFP images of an early-stage embryo carrying P csp-3 csp-3::gfp are shown.

    Journal: Nature structural & molecular biology

    Article Title: Inhibition of CED-3 zymogen activation and apoptosis in Caenorhabditis elegans by caspase homolog CSP-3

    doi: 10.1038/nsmb.1488

    Figure Lengend Snippet: CSP-3 is a cytoplasmic protein with sequence homology to the small subunit of CED-3 ( a ) Sequence alignment of CSP-3 and the small subunits of CED-3 and human caspase-3. Residues that are identical are shaded in yellow and residues that are similar are labeled with pink. Phe57 of CSP-3 is indicated with a blue arrowhead. ( b ) The csp-3 gene structure and deletion mutations. Exons are depicted as boxes and introns as lines. The translated regions of csp-3 are highlighted in blue. Two orange boxes indicate the regions of csp-3 removed by the two csp-3 deletions, respectively. Also shown are a csp-3 translational gfp fusion and a 5.73-kb genomic rescuing fragment ( csp-3 (+)). ( c ) Both csp-3 deletions abolish expression of csp-3 in C. elegans . Immunoblotting for CSP-3 in total worm lysates from N2 , csp-3 ( tm2260 ) and csp-3 ( tm2486 ) animals is shown. The CstF-64 protein was used as a loading control. ( d ) csp-3 is ubiquitously expressed in C. elegans embryos. Differential interference contrast (DIC) and GFP images of an early-stage embryo carrying P csp-3 csp-3::gfp are shown.

    Article Snippet: In vitro CED-3 zymogen activation assay The CED-3 zymogen was first synthesized and labeled with 35 S-methionine in the TNT Transcription/Translation coupled system (Promega) at 30 °C as described previously , in the presence of an equal amount of GST–CSP-3, GST–CSP-3(F57D) or GST.

    Techniques: Sequencing, Labeling, Expressing

    CSP-3 associates with CED-3 in vitro and in C. elegans ( a ) CSP-3 binds to the CED-3 zymogen. GST–CSP-3, GST–CSP-3(F57D) or GST was coexpressed in bacteria with the CED-3 zymogen tagged with a Flag epitode (CED-3–Flag). One portion of the soluble fraction was analyzed by western blot (IB) to examine the expression levels of GST fusion proteins and CED-3–Flag. The remaining portion of the soluble fraction was used for GST protein pull-down experiment, and the amount of CED-3–Flag pulled down was analyzed by western blot analysis. ( b ) CSP-3 associates specifically with the large subunit of CED-3 in vitro . GST–CSP-3, GST–CSP-3(F57D) or GST was coexpressed in bacteria with the CED-3 large subunit (p17) or the small subunit (p13), both tagged with a Flag epitode (gray box). Analysis of expression levels as well as the amounts of two CED-3 subunits coprecipitated with GST fusion proteins was conducted as described in a . The diagram above depicts the domain structure of the CED-3 zymogen, with arrows indicating the three proteolytic cleavage sites that lead to the activation of the CED-3 zymogen. The three CED-3 cleavage products are shown below as boxes. ( c ) CSP-3 associates with CED-3 in C. elegans . Lysates from C. elegans animals expressing CED-3::GFP or CEH-30::GFP were prepared as described in Methods . One portion of the worm lysate was used in the western blot analysis to examine the expression levels of CSP-3 and GFP fusion proteins. The remaining portion of the lysate was incubated with a mouse anti-GFP monoclonal antibody and precipitated using Protein G Sepharose beads. The amount of the CSP-3 protein pulled down with the GFP fusion proteins was analyzed by western blot using purified anti–CSP-3 antibody. A small amount of full-length CED-3::GFP fusion was detected in the lysate (data not shown). The predominant species detected was CED-3::GFP fusion without its prodomain but containing both large and small subunits.

    Journal: Nature structural & molecular biology

    Article Title: Inhibition of CED-3 zymogen activation and apoptosis in Caenorhabditis elegans by caspase homolog CSP-3

    doi: 10.1038/nsmb.1488

    Figure Lengend Snippet: CSP-3 associates with CED-3 in vitro and in C. elegans ( a ) CSP-3 binds to the CED-3 zymogen. GST–CSP-3, GST–CSP-3(F57D) or GST was coexpressed in bacteria with the CED-3 zymogen tagged with a Flag epitode (CED-3–Flag). One portion of the soluble fraction was analyzed by western blot (IB) to examine the expression levels of GST fusion proteins and CED-3–Flag. The remaining portion of the soluble fraction was used for GST protein pull-down experiment, and the amount of CED-3–Flag pulled down was analyzed by western blot analysis. ( b ) CSP-3 associates specifically with the large subunit of CED-3 in vitro . GST–CSP-3, GST–CSP-3(F57D) or GST was coexpressed in bacteria with the CED-3 large subunit (p17) or the small subunit (p13), both tagged with a Flag epitode (gray box). Analysis of expression levels as well as the amounts of two CED-3 subunits coprecipitated with GST fusion proteins was conducted as described in a . The diagram above depicts the domain structure of the CED-3 zymogen, with arrows indicating the three proteolytic cleavage sites that lead to the activation of the CED-3 zymogen. The three CED-3 cleavage products are shown below as boxes. ( c ) CSP-3 associates with CED-3 in C. elegans . Lysates from C. elegans animals expressing CED-3::GFP or CEH-30::GFP were prepared as described in Methods . One portion of the worm lysate was used in the western blot analysis to examine the expression levels of CSP-3 and GFP fusion proteins. The remaining portion of the lysate was incubated with a mouse anti-GFP monoclonal antibody and precipitated using Protein G Sepharose beads. The amount of the CSP-3 protein pulled down with the GFP fusion proteins was analyzed by western blot using purified anti–CSP-3 antibody. A small amount of full-length CED-3::GFP fusion was detected in the lysate (data not shown). The predominant species detected was CED-3::GFP fusion without its prodomain but containing both large and small subunits.

    Article Snippet: In vitro CED-3 zymogen activation assay The CED-3 zymogen was first synthesized and labeled with 35 S-methionine in the TNT Transcription/Translation coupled system (Promega) at 30 °C as described previously , in the presence of an equal amount of GST–CSP-3, GST–CSP-3(F57D) or GST.

    Techniques: In Vitro, Western Blot, Expressing, Activation Assay, Incubation, Purification

    Role of autophagy induction on the inhibition of apoptosis by globular adiponectin in HepG2 cells. (A) Cells were transfected with LC3B siRNA or scrambled control siRNA followed by incubation with gAcrp (1 μg/ml) for 48 h. Cell viability was assessed as described previously. (B) Cells were transfected with LC3B siRNA or scrambled siRNA followed by gAcrp (1 μg/ml) incubation for 48 h. Caspase-3 activity was determined as described previously. (C) Cells were pretreated with indicated concentration of rapamycin for 2 h followed by treatment with gAcrp (1 μg/ml) for 48 h. Caspase-3 activity was determined as described previously. Values represent fold increase compared to control and expressed as mean ± SEM (A and C; n=3: B; n=4). * p

    Journal: Biomolecules & Therapeutics

    Article Title: Regulatory Role of Autophagy in Globular Adiponectin-Induced Apoptosis in Cancer Cells

    doi: 10.4062/biomolther.2014.021

    Figure Lengend Snippet: Role of autophagy induction on the inhibition of apoptosis by globular adiponectin in HepG2 cells. (A) Cells were transfected with LC3B siRNA or scrambled control siRNA followed by incubation with gAcrp (1 μg/ml) for 48 h. Cell viability was assessed as described previously. (B) Cells were transfected with LC3B siRNA or scrambled siRNA followed by gAcrp (1 μg/ml) incubation for 48 h. Caspase-3 activity was determined as described previously. (C) Cells were pretreated with indicated concentration of rapamycin for 2 h followed by treatment with gAcrp (1 μg/ml) for 48 h. Caspase-3 activity was determined as described previously. Values represent fold increase compared to control and expressed as mean ± SEM (A and C; n=3: B; n=4). * p

    Article Snippet: Caspase-3 activity assay kit and cell proliferation assay kit (MTS) were purchased from Promega Corporation (Madison, WI, USA).

    Techniques: Inhibition, Transfection, Incubation, Activity Assay, Concentration Assay

    Globular adiponectin induces apoptosis in HepG2 cells. Cells were treated with gAcrp (1 μg/ml) for the indicated time points. Cell viability (A) and caspase-3 activity (B) were assessed by MTS assay and Caspase-3 activity assay, respectively. Values represent percentage of cell death or fold increase compared to control and expressed as mean ± SEM (n=3). * p

    Journal: Biomolecules & Therapeutics

    Article Title: Regulatory Role of Autophagy in Globular Adiponectin-Induced Apoptosis in Cancer Cells

    doi: 10.4062/biomolther.2014.021

    Figure Lengend Snippet: Globular adiponectin induces apoptosis in HepG2 cells. Cells were treated with gAcrp (1 μg/ml) for the indicated time points. Cell viability (A) and caspase-3 activity (B) were assessed by MTS assay and Caspase-3 activity assay, respectively. Values represent percentage of cell death or fold increase compared to control and expressed as mean ± SEM (n=3). * p

    Article Snippet: Caspase-3 activity assay kit and cell proliferation assay kit (MTS) were purchased from Promega Corporation (Madison, WI, USA).

    Techniques: Activity Assay, MTS Assay, Caspase-3 Activity Assay

    PHGDH-derived serine supports the transsulfuration and folate cycles. (a) Serine metabolism via the transsulfuration and folate cycles. 13 C-labelled carbons (●) unlabelled carbons ( ○ ). Gly: Glycine, Ser: Serine, CTH: Cystathionine; HCY: Homocysteine, GSH: Glutathione, γGC: γ-glutamyl cysteine, Glu: Glutamate. (b–e) A549 cells expressing scramble (SCR), NRF2 or PHGDH shRNAs were grown in the presence of U- 13 C-glucose for 24 hours and metabolites were extracted and analysed by LC/MS. (b) Analysis of 13 C-labelling on the glycine component of glutathione. (c) Analysis of 13 C-labeling on the purine metabolite adenosine monophosphate (AMP). (d,e) Analysis of glutathione (d) and AMP (e) labelling in serine high cell lines. Cells were labelled with 13 C-glucose for 24 or 48 hours as indicated. (b,d) PHGDH-derived serine is incorporated into glutathione through the generation of glycine and cysteine. M+0 denotes no carbons labelled, M+2 denotes labelling on the glycine moiety, M+3 denotes labeling on the cysteine moiety, and M+5 denotes labeling on both glycine and cysteine. M+5 labeling was not observed. (c,e) M+5 labelling occurs following ribose-5-phosphate labelling via the pentose phosphate pathway. M+7 labelling is the result of ribose labelling plus either glycine or formyl-THF labelling in the purine ring. M+9 labelling is the result of ribose labelling plus either glycine and formyl-THF labelling in the purine ring. M+0 has no labelled carbons. (f–g) LC/MS analysis of total metabolite levels in the nucleotide (f) and transsulfuration (g) pathways. (h) NADPH/NADP+ ratios 4 days after PHGDH knockdown. Results are the average of 3 biological replicates.

    Journal: Nature genetics

    Article Title: NRF2 regulates serine biosynthesis in non-small cell lung cancer

    doi: 10.1038/ng.3421

    Figure Lengend Snippet: PHGDH-derived serine supports the transsulfuration and folate cycles. (a) Serine metabolism via the transsulfuration and folate cycles. 13 C-labelled carbons (●) unlabelled carbons ( ○ ). Gly: Glycine, Ser: Serine, CTH: Cystathionine; HCY: Homocysteine, GSH: Glutathione, γGC: γ-glutamyl cysteine, Glu: Glutamate. (b–e) A549 cells expressing scramble (SCR), NRF2 or PHGDH shRNAs were grown in the presence of U- 13 C-glucose for 24 hours and metabolites were extracted and analysed by LC/MS. (b) Analysis of 13 C-labelling on the glycine component of glutathione. (c) Analysis of 13 C-labeling on the purine metabolite adenosine monophosphate (AMP). (d,e) Analysis of glutathione (d) and AMP (e) labelling in serine high cell lines. Cells were labelled with 13 C-glucose for 24 or 48 hours as indicated. (b,d) PHGDH-derived serine is incorporated into glutathione through the generation of glycine and cysteine. M+0 denotes no carbons labelled, M+2 denotes labelling on the glycine moiety, M+3 denotes labeling on the cysteine moiety, and M+5 denotes labeling on both glycine and cysteine. M+5 labeling was not observed. (c,e) M+5 labelling occurs following ribose-5-phosphate labelling via the pentose phosphate pathway. M+7 labelling is the result of ribose labelling plus either glycine or formyl-THF labelling in the purine ring. M+9 labelling is the result of ribose labelling plus either glycine and formyl-THF labelling in the purine ring. M+0 has no labelled carbons. (f–g) LC/MS analysis of total metabolite levels in the nucleotide (f) and transsulfuration (g) pathways. (h) NADPH/NADP+ ratios 4 days after PHGDH knockdown. Results are the average of 3 biological replicates.

    Article Snippet: NADPH/NADP+ ratios NADP+ and NADPH were determined with the NADP/NADPH-glo assay kit (Promega) according to the manufacturer’s protocol.

    Techniques: Derivative Assay, Expressing, Liquid Chromatography with Mass Spectroscopy, Labeling