The Experts below are selected from a list of 213 Experts worldwide ranked by ideXlab platform
Rolf W Hartmann - One of the best experts on this subject based on the ideXlab platform.
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5α reductase in human embryonic Kidney Cell Line hek293 evidence for type ii enzyme expression and activity
Molecular and Cellular Biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5α-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5α-reductase type II activity. After an incubation of 24 h 1 × 106 HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5α-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5α-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5α-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5α-reductase type II protein. (Mol Cell Biochem 270: 201–208, 2005)
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5alpha-reductase in human embryonic Kidney Cell Line HEK293: evidence for type II enzyme expression and activity.
Molecular and cellular biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5alpha-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5alpha-reductase type II activity. After an incubation of 24 h 1 x 10(6) HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5alpha-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5alpha-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5alpha-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5alpha-reductase type II protein.
Baerbel U Panter - One of the best experts on this subject based on the ideXlab platform.
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5α reductase in human embryonic Kidney Cell Line hek293 evidence for type ii enzyme expression and activity
Molecular and Cellular Biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5α-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5α-reductase type II activity. After an incubation of 24 h 1 × 106 HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5α-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5α-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5α-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5α-reductase type II protein. (Mol Cell Biochem 270: 201–208, 2005)
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5alpha-reductase in human embryonic Kidney Cell Line HEK293: evidence for type II enzyme expression and activity.
Molecular and cellular biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5alpha-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5alpha-reductase type II activity. After an incubation of 24 h 1 x 10(6) HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5alpha-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5alpha-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5alpha-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5alpha-reductase type II protein.
Joachim Jose - One of the best experts on this subject based on the ideXlab platform.
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5α reductase in human embryonic Kidney Cell Line hek293 evidence for type ii enzyme expression and activity
Molecular and Cellular Biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5α-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5α-reductase type II activity. After an incubation of 24 h 1 × 106 HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5α-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5α-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5α-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5α-reductase type II protein. (Mol Cell Biochem 270: 201–208, 2005)
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5alpha-reductase in human embryonic Kidney Cell Line HEK293: evidence for type II enzyme expression and activity.
Molecular and cellular biochemistry, 2005Co-Authors: Baerbel U Panter, Joachim Jose, Rolf W HartmannAbstract:Human 5alpha-reductase catalyses the last step in androgen biosynthesis, namely the reduction of testosterone (T) to the more potent androgen dihydrotestosterone (DHT). The enzyme is therefore considered to be an important drug target for androgen related diseases such as benign prostatic hyperplasia and prostate cancer. The present study displays evidence that the human embryonic Kidney Cell Line HEK293 which is frequently used in recombinant target protein expression contains an endogenous 5alpha-reductase type II activity. After an incubation of 24 h 1 x 10(6) HEK293 Cells converted 23% of the substrate 4-androstene-3,17-dione (7.5 nM) to the product 5alpha-androstane-3,17-dione. Reverse transcription polymerase chain reaction was carried out to identify the mRNA of the isoform responsible for the 5alpha-reductase activity. Only with type II specific primers a fragment with the predicted size was amplified, while with type I specific primers no band could be observed. An antiserum against human 5alpha-reductase type II was raised by immunizing a rabbit with a hemocyanin-conjugated peptide corresponding to amino acid 29 to 44 of the type II enzyme. Western blot analysis of different fractions of a HEK293 homogenate performed with this antiserum detected a band at 45 kDa in the nuclear and microsomal fraction corresponding to 5alpha-reductase type II protein.
Luis L Rodriguez - One of the best experts on this subject based on the ideXlab platform.
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A continuous bovine Kidney Cell Line constitutively expressing bovine αvβ6 integrin has increased susceptibility to foot-and-mouth disease virus.
Journal of clinical microbiology, 2013Co-Authors: Michael Larocco, Peter W Krug, Ed Kramer, Zaheer Ahmed, Juan M Pacheco, Hernando Duque, Barry Baxt, Luis L RodriguezAbstract:Foot-and-mouth disease (FMD) is a worldwide problem limiting the trade of animals and their products from affected countries. The rapid isolation, serotyping, and vaccine matching of FMD virus from disease outbreaks is critical for enabling the implementation of effective vaccination programs and to stop the spread of infection during outbreaks. Some primary Cells have been shown to be highly susceptible to most strains of FMD virus (FMDV) but are difficult and expensive to prepare and maintain. Since the αVβ6 integrin is a principal receptor for FMDV, we transduced a bovine Kidney Cell Line to stably express both the αV and β6 bovine integrin subunits. This stable Cell Line (LFBK-αVβ6) showed β6 expression and enhanced susceptibility to FMDV infection for ≥ 100 Cell passages. LFBK-αVβ6 Cells were highly sensitive for detecting all serotypes of FMDV from experimentally infected animals, including the porcinophilic FMDV strain O/TAW/97. In comparison to other Cell types that are currently used for virus isolation, LFBK-αVβ6 Cells were more effective at detecting FMDV in clinical samples, supporting their use as a more sensitive tool for virus isolation.
Peter G.w. Plagemann - One of the best experts on this subject based on the ideXlab platform.
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High-affinity, equilibrative nucleoside transporter of pig Kidney Cell Line (PK-15)
Biochimica et biophysica acta, 1992Co-Authors: Josep M. Aran, Peter G.w. PlagemannAbstract:Abstract Nucleoside transport was determined in the cloned porcine Kidney Cell Line PK-15 which exhibits properties of tubular Cells. The Cells did not express any Na+-dependent, concentrative nucleoside transport. They exhibited only nitrobenzylthioinosine-sensitive equilibrative nucleoside transport. Their transport activity, however, was only about 10% of that observed in many other mammalian Cell Lines. This low transport activity correlated with a relatively low number of high-affinity nitrobenzylthioinosine binding sites (5·103 /Cell). Furthermore, although the equilibrative transporter of PK-15 Cells exhibited a similar broad substrate specificity as the equilibrative nucleoside transporters of other mammalian Cells, it exhibited a much higher affinity for certain nucleosides, especially cytidine and uridine, than the latter. The Michaelis-Menten constants for zero-trans transport and equilibrium exchange of uridine in ATP-depleted Cells were about the same (about 40 μM), indicating equal mobility of the nucleoside-loaded and empty carrier. Concentrative nucleoside transport was detected in one set of PK-15 cultures, but was found to be due to mycoplasma contamination.
