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    PerkinElmer perkinelmer wizard2 γ counter
    Perkinelmer Wizard2 γ Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 76/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer 2480 wizard2 gamma counter perkinelmer
    Generation and characterization of Titanium-45. (A) A <t>gamma</t> spectrum of 45 Ti obtained with the HPGe detector. (B) and (C) show the enlarged spectra of two characteristic gammas from 45 Ti at 720 and 1408 keV (pointed by link and blue arrows). Inset in (A) shows the MicroPET image of a Derenzo phantom of 45 Ti. The distance values shown in the phantom image are the diameters of the cylinders in each region. The distance between axes of the cylinders is twice their diameter.
    2480 Wizard2 Gamma Counter Perkinelmer, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 80/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer perkinelmer gamma counter
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    Perkinelmer Gamma Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 80/100, based on 309 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer perkinelmer γ counter
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    Perkinelmer γ Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 85/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer γ 32 p atp nen perkinelmer
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    γ 32 P Atp Nen Perkinelmer, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer auto gamma 5530 minaxi γ counter
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    Auto Gamma 5530 Minaxi γ Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer γ 32 p atp perkinelmer waltham ma
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    γ 32 P Atp Perkinelmer Waltham Ma, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer γ counter wallac 1470 wizard gamma counter
    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and <t>gamma</t> counting.
    γ Counter Wallac 1470 Wizard Gamma Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 83/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer atp γ 32p easytide
    H2A monoubiquitination assay and CK2 kinase assay performed with γ- 32 <t>P-ATP.</t> a Coomassie blue staining of factors used. b Scheme for H2A monoubiquitination assay with E1 that was pre-charged with HA-ubiquitin (See Methods for details). c Immunoblotting of H2A monoubiquitination assay as describe in b with increasing amounts of CK2. d Radiograph of CK2 kinase assay reaction products. The assay was assembled with the factors indicated, each at the same amount used in the H2A monoubiquitination assay (Methods). After incubation at 37°C for 30 min, the assay was stopped by boiling in SDS loading buffer and resolved on SDS-PAGE. Besides CK2B, which was radio-labeled presumably due to autophosphorylation, phosphorylation of RING1B and PCGF5 was detected together with a species, indicated as *histone, dependent on the presence of nucleosomes. e H2A monoubquitination assay performed as in Fig. 3c , using increasing amounts of RING1B-PCGF5-AUTS2 containing either RING1B-S41A (S41 to alanine), or RING1B-S41D (S41 to aspartic acid), purified from sf9 cells.
    Atp γ 32p Easytide, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 90/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer atp γ 32p easytide lead
    Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 <t>P]ATP.</t> The result of the reaction was run through an SDS-polyacrylamide
    Atp γ 32p Easytide Lead, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 90/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer cobra ii auto gamma γ counter
    Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 <t>P]ATP.</t> The result of the reaction was run through an SDS-polyacrylamide
    Cobra Ii Auto Gamma γ Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 78/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma atp33
    Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 <t>P]ATP.</t> The result of the reaction was run through an SDS-polyacrylamide
    Gamma Atp33, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 81/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma 32p
    Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 <t>P]ATP.</t> The result of the reaction was run through an SDS-polyacrylamide
    Gamma 32p, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer γ p32
    Action of CK1δ within the purified nuclear PER complex (A) Protein kinase activity of nuclear PER complex. BN-PAGE autoradiogram showing control sample (WT) or affinity-purified nuclear PER complex (PER2-FH) after incubation with γ- 32 P-ATP (25°C, 1 h). . (C) CK1δ in purified PER complex phosphorylates PERs and CLOCK. SDS-PAGE autoradiogram as in (B). Leftmost lane, no inhibitor. Remaining lanes, decreasing concentrations (625, 125, 25, or 5 nM, represented by wedges) of PF-670462, a CK1δ/ε inhibitor (CK1δ/ε), or PF-4800567, a CK1ε-selective inhibitor (CK1ε). (D) Top and Middle, BN-APAGE immunoblots of mouse liver nuclear extracts (CT18) from wildtype (WT) or single mutants null for the circadian clock genes indicated at the top (KO) probed for proteins indicated at right. Bottom, SDS-PAGE SAP155 immunoblot loading control. (E) Top, SDS-PAGE autoradiogram of liver nuclear extracts (CT18) from genotypes indicated at top after immunoprecipitation with anti-CRY1 antibody and incubation with γ- 32 P-ATP (25°C, 10 min; linear range for wildtype). Bottom, SDS-PAGE immunoblot for CLOCK served as loading control for PER complexes. (F) Phosphor-imaging quantification of labeled CLOCK bands from (E) after normalization to CLOCK immunoblot signals from (E). Shown are mean ± SEM;  t -test (two-tailed). Dashed line, background.
    γ P32, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 81/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer atp
    Reconstitution and 3D reconstruction of the PI3KC3-C1 complex. ( A ) Domain structures of the four subunits of PI3KC3-C1. The small unlabeled yellow box between VPS34 C2 and helical domains is the CHIL motif, described below. ( B ) Purification of PI3KC3-C1. Coomassie-stained SDS-PAGE gel of purified PI3KC3-C1. ( C ) Thin-layer chromatography of radiolabeled PI(3)P generated by PI3KC3-C1 from PI and <t>[γ-32</t> P] <t>ATP.</t> ( D ) Reference-free class averages of PI3KC3-C1, each containing ∼200 particles. The first row represents particles that were selected for the 3D reconstruction. The lower two rows represent particles excluded from the 3D reconstruction, the arrowhead indicating the position of a density dislodged from the main part of the complex. All class averages shown are calculated from data acquired on untilted grids, except the top right class average which is from 45° tilt. ( E ) 3D reconstruction calculated from ∼39,000 particles in four orientations and displayed at a threshold determined from subsequent docking analyses. DOI: http://dx.doi.org/10.7554/eLife.05115.003
    Atp, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 90/100, based on 669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gtp
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Gtp, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 85/100, based on 101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma labeled
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Gamma Labeled, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer wizard2 gamma counter
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Wizard2 Gamma Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 98/100, based on 260 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma counter 2480wizard2
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Gamma Counter 2480wizard2, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma type counter
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Gamma Type Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer ls1500 gamma counter
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
    Ls1500 Gamma Counter, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 77/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma alpha counter
    PI5P4Kβ has ability to sense <t>GTP</t> concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM <t>γ-</t> 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .
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    PerkinElmer scintillation gamma counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    PerkinElmer wallace gamma counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    PerkinElmer wallac gamma counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    PerkinElmer gamma counter a 2480 automatic gamma counter wizard2 3
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
    Gamma Counter A 2480 Automatic Gamma Counter Wizard2 3, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer topcount gamma counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    PerkinElmer riastar gamma counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    PerkinElmer gamma spectroscopy
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
    Gamma Spectroscopy, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 89/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer gamma scintillator counter
    Representative <t>gamma</t> images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.
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    94
    PerkinElmer ifn γ
    Immunomodulatory effect of endogenous IL-18 and IL-12 on intrapulmonary <t>IFN-γ</t> during replicative L. pneumophila lung infection. A/J mice were administered anti-IL-18R MAb, anti-IL-12 MAb, or both MAbs prior to intratracheal inoculation with L. pneumophila . At specific time points p.i., mice were euthanatized, and lungs were excised, homogenized, and filtered. Intrapulmonary levels of IFN-γ were quantified in filtered lung homogenates by ELISA. Mice were treated with control antibody (IgG2a) (solid bars), anti-IL-12 MAb (shaded bars), anti-IL-18RmAB (hatched bars), anti-IL-12 MAb and anti-IL-18R MAb (horizontally striped bars). Results represent means ± standard errors of the means for five animals per treatment group. ∗, P
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    PerkinElmer atp γ 32p lead
    Cdk1 is important for ESC pluripotency and preferentially interacts with Oct4 at the G2/M phase. ( A ) Protein levels and mRNA levels of Cdk1-knockdown mESCs ( n = 3). ( B and C ) AP staining of Cdk1-knockdown mESCs. After staining to reveal AP activity, the colonies were scored and the percentages of undifferentiated, partially differentiated, and fully differentiated colonies were calculated. AP staining experiments were repeated three times ( n = 3). Scale bar represents 500 μm. ( D ) Immunostaining of Cdk1-knockdown mESCs. Cdk1 was stained with anti-Cdk1 (green) and DNA was stained DAPI (blue). Scale bar represents 100 μm. ( E ) Fluorescence images of Cdk1-knockdown mESCs. Nanog and Klf4 were stained with anti-Nanog (green) and anti-Klf4 (green), respectively. DNA was stained with DAPI (blue). Scale bar represents 100 μm. ( F ) Flag-Oct4 and HA-Cdk1 were cotransfected into HEK293T cells. Cell lysates were immunoprecipitated with anti-Flag antibody and probed with anti-HA antibody (left). Endogenous Cdk1 was immunoprecipitated from E14 mESCs with anti-Oct4 antibody and immunoblotted with anti-Cdk1 antibody (right). ( G ) Changes in interaction of Cdk1 with Oct4 during cell cycle progression. Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole (100 ng/ml) for 8 h. At the indicated time after release into fresh media, cell cycle progression was determined by FACS analysis (right). Cell lysates were pulled down with anti-Flag beads. Bound proteins were eluted and then immunoblotted with the indicated antibodies. (A, asynchronous state; N, nocodazole treatment). ( H ) Changes in interaction of Oct4 with Cdk1 depending on Cyclin B knockdown. After Cyclin B knockdown, Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole for 8 h. Flag-Oct4 was immunoprecipitated by incubating lysates with anti-Flag beads. Following Flag elution, bound proteins were immunoblotted with the indicated antibodies. ( I ) Colocalization of Cdk1 and p-Oct4(S229) in mitosis-arrested mESCs was analyzed by immunostaining. mESCs were treated with nocodazole (100 ng/ml) for 8 h. Cdk1 was stained with anti-Cdk1 (green), p-Oct4(S229) was stained with anti-p-Oct4(S229) (red), and DNA was stained with DAPI (blue). Scale bar represents 5 μm. ( J ) Radioactive in vitro kinase assay using recombinant Cdk1 to phosphorylate GST-Oct4 wild type (WT) and S228A/S229A mutant. Autoradiogram showing incorporation of γ- 32 P <t>ATP</t> (left). For the IP kinase assay, cell lysates of nocodazole-treated E14 mESCs were immunoprecipitated with anti-CycB1 and Aurkb and subjected to an in vitro kinase assay using (His) 6 -PP1α and GST-Oct4 as a substrate and followed by western blotting (right).
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    Image Search Results


    Generation and characterization of Titanium-45. (A) A gamma spectrum of 45 Ti obtained with the HPGe detector. (B) and (C) show the enlarged spectra of two characteristic gammas from 45 Ti at 720 and 1408 keV (pointed by link and blue arrows). Inset in (A) shows the MicroPET image of a Derenzo phantom of 45 Ti. The distance values shown in the phantom image are the diameters of the cylinders in each region. The distance between axes of the cylinders is twice their diameter.

    Journal: Acta Pharmacologica Sinica

    Article Title: Intrinsic radiolabeling of Titanium-45 using mesoporous silica nanoparticles

    doi: 10.1038/aps.2017.1

    Figure Lengend Snippet: Generation and characterization of Titanium-45. (A) A gamma spectrum of 45 Ti obtained with the HPGe detector. (B) and (C) show the enlarged spectra of two characteristic gammas from 45 Ti at 720 and 1408 keV (pointed by link and blue arrows). Inset in (A) shows the MicroPET image of a Derenzo phantom of 45 Ti. The distance values shown in the phantom image are the diameters of the cylinders in each region. The distance between axes of the cylinders is twice their diameter.

    Article Snippet: The radioactivity in the tissue was measured using a gamma-counter (Perkin-Elmer) and presented as%ID/g (%ID/g=percentage injected dose per gram, mean±SD).

    Techniques:

    A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and gamma counting.

    Journal: Theranostics

    Article Title: Evaluation of an [18F]AlF-NOTA Analog of Exendin-4 for Imaging of GLP-1 Receptor in Insulinoma

    doi: 10.7150/thno.5276

    Figure Lengend Snippet: A) Representative PET images of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 in INS-1 xenografted mice at 30 and 60 min p.i.. Tumor uptake is completely blocked by co-administration of a blocking dose of exendin-4. B) Quantification of uptake by selected tissues determined by ROIs following injection of [ 18 F]AlF-NOTA-MAL-cys 40 -exendin-4 or co-injection of blocking agent (exendin-4) into INS-1 tumor-bearing mice at 30 and 60 min p.i.. C) Experiment of B at 60 min with analysis by tissue dissection and gamma counting.

    Article Snippet: Radioactivity was determined using an on-line radioactivity detector or collection of 1-min fractions followed by gamma-counting (Wallach Wizard, PerkinElmer, Waltham, MA).

    Techniques: Positron Emission Tomography, Mouse Assay, Blocking Assay, Injection, Dissection

    Tf supplementation simulates iron release from iron-replete cells and protects against MPTP in cells and mice. ( a ) Iron release from 59 Fe-loaded M17 cells was measured by γ-counter. ( b ) M17 cells were treated with MPP+ in the absence and presence of apo-Tf for 24 h. Cell viability assessed using the MTT assay. ( c , d ) MPTP (50 mg kg −1 ) was administered to mice. A subgroup of mice was subsequently administered TF (30 mg kg −1 ; intraperitoneal) every third day for 21 days and compared with controls ( N =9 each). ( c ) Mice performed the pole test 20 days after MPTP lesion. ( d ) Iron (AAS) content in nigra of MPTP and Tf-treated mice expressed as fold elevation of controls. Data are means±s.e.m. * P

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Transferrin protects against Parkinsonian neurotoxicity and is deficient in Parkinson’s substantia nigra

    doi: 10.1038/sigtrans.2016.15

    Figure Lengend Snippet: Tf supplementation simulates iron release from iron-replete cells and protects against MPTP in cells and mice. ( a ) Iron release from 59 Fe-loaded M17 cells was measured by γ-counter. ( b ) M17 cells were treated with MPP+ in the absence and presence of apo-Tf for 24 h. Cell viability assessed using the MTT assay. ( c , d ) MPTP (50 mg kg −1 ) was administered to mice. A subgroup of mice was subsequently administered TF (30 mg kg −1 ; intraperitoneal) every third day for 21 days and compared with controls ( N =9 each). ( c ) Mice performed the pole test 20 days after MPTP lesion. ( d ) Iron (AAS) content in nigra of MPTP and Tf-treated mice expressed as fold elevation of controls. Data are means±s.e.m. * P

    Article Snippet: Media was collected after 3 h and measured for 59 Fe release by γ-counter (Wizard 3, PerkinElmer) and expressed as counts per min.

    Techniques: Mouse Assay, MTT Assay, Atomic Absorption Spectroscopy

    Spatial accuracy and planar imaging. ( A ) CT images acquired postmilling from six mice with three regions each. The VOIs, as defined before milling, are circled in green. ( B and C ) H E stains of IGMM-derived tissue samples from a VOI placed in the heart ( B ) and from a VOI placed in the kidney ( C ). ( D – G ) A regular light photograph (adjusted for white-balance and slight geometric distortions) ( D ), CT ( E ), PET ( F ), and OI ( G ) in a plane through a mouse. D and G were obtained after milling. ( H ) Activity concentrations obtained with PET and a gamma-counter in 11 cylinders.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Linking imaging to omics utilizing image-guided tissue extraction

    doi: 10.1073/pnas.1718304115

    Figure Lengend Snippet: Spatial accuracy and planar imaging. ( A ) CT images acquired postmilling from six mice with three regions each. The VOIs, as defined before milling, are circled in green. ( B and C ) H E stains of IGMM-derived tissue samples from a VOI placed in the heart ( B ) and from a VOI placed in the kidney ( C ). ( D – G ) A regular light photograph (adjusted for white-balance and slight geometric distortions) ( D ), CT ( E ), PET ( F ), and OI ( G ) in a plane through a mouse. D and G were obtained after milling. ( H ) Activity concentrations obtained with PET and a gamma-counter in 11 cylinders.

    Article Snippet: For comparison of PET and gamma-counter uptake, the samples were weighed and transferred to an automatic gamma-counter (WIZARD2 2480; PerkinElmer) for determination of radioactivity and calculation of the activity concentration.

    Techniques: Imaging, Mouse Assay, Derivative Assay, Positron Emission Tomography, Activity Assay

    Typical representative time-activity curve from an ip injection of 18 F-FDG. Errors from gamma counting statistics (

    Journal: American Journal of Nuclear Medicine and Molecular Imaging

    Article Title: An anesthetic method compatible with 18F-FDG-PET studies in mice

    doi:

    Figure Lengend Snippet: Typical representative time-activity curve from an ip injection of 18 F-FDG. Errors from gamma counting statistics (

    Article Snippet: Whole blood 18 F-FDG concentration was measured in an automated gamma-well counter (1480 Perkin Elmer, Wizard 3).

    Techniques: Activity Assay, Injection

    The biodistribution of some organs and tumors in mice The γ-counter quantification analysis for uptakes of some organs and tumors for 18 F-FDG in H1975 xenograft models (n=10) after dynamic imaging 60min.

    Journal: Oncotarget

    Article Title: High-resolution dynamic imaging and quantitative analysis of lung cancer xenografts in nude mice using clinical PET/CT

    doi: 10.18632/oncotarget.17263

    Figure Lengend Snippet: The biodistribution of some organs and tumors in mice The γ-counter quantification analysis for uptakes of some organs and tumors for 18 F-FDG in H1975 xenograft models (n=10) after dynamic imaging 60min.

    Article Snippet: The radioactivity of wet tissues was counted for 30s in an automatic γ-counter (WIZARD 2480, Perkin Elmer, New York, USA) [ ].

    Techniques: Mouse Assay, Imaging

    Linear regression analysis of %ID/g from clinical PET/CT versus γ-counter The %ID/g of main tissues extracted from PET/CT with SharpIR reconstruction and that by ex vivo counting has a strong correlation (r 2 = 0.995). A marked improvement of the slope (0.981) when data was reconstructed with SharpIR reconstruction algorithm compared to without SharpIR (0.634).

    Journal: Oncotarget

    Article Title: High-resolution dynamic imaging and quantitative analysis of lung cancer xenografts in nude mice using clinical PET/CT

    doi: 10.18632/oncotarget.17263

    Figure Lengend Snippet: Linear regression analysis of %ID/g from clinical PET/CT versus γ-counter The %ID/g of main tissues extracted from PET/CT with SharpIR reconstruction and that by ex vivo counting has a strong correlation (r 2 = 0.995). A marked improvement of the slope (0.981) when data was reconstructed with SharpIR reconstruction algorithm compared to without SharpIR (0.634).

    Article Snippet: The radioactivity of wet tissues was counted for 30s in an automatic γ-counter (WIZARD 2480, Perkin Elmer, New York, USA) [ ].

    Techniques: Positron Emission Tomography, Ex Vivo

    Regression plots for PET/CT SUV max versus %ID/g from γ-counter in main tissues There was a low correlation (r 2 = 0.745, slope = 0.116; P

    Journal: Oncotarget

    Article Title: High-resolution dynamic imaging and quantitative analysis of lung cancer xenografts in nude mice using clinical PET/CT

    doi: 10.18632/oncotarget.17263

    Figure Lengend Snippet: Regression plots for PET/CT SUV max versus %ID/g from γ-counter in main tissues There was a low correlation (r 2 = 0.745, slope = 0.116; P

    Article Snippet: The radioactivity of wet tissues was counted for 30s in an automatic γ-counter (WIZARD 2480, Perkin Elmer, New York, USA) [ ].

    Techniques: Positron Emission Tomography

    Silencing of NALP3 in THP1 cells abolishes protein permeability across mouse ATII cell monolayers in ATII and THP1 coculture models. A , ATII cells were grown in 24-well transwell plates until the cellular monolayers reached confluence. THP-1 cells were added to the appropriate ATII monolayers, cocultured for 6 h, and exposed to hyperoxic conditions for 1 h. In some experiments, THP-1 cells were transfected with control shRNA or NALP3 shRNA and then added to a monolayer of ATII cells. Transepithelial albumin flux across each monolayer was determined by the addition of [ 125 I]-labeled human serum albumin to each upper compartment. The contents from the lower compartment were collected and counted in a Wizard γ-counter. * p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: The Inflammasome Mediates Hyperoxia-Induced Alveolar Cell Permeability

    doi: 10.4049/jimmunol.0902766

    Figure Lengend Snippet: Silencing of NALP3 in THP1 cells abolishes protein permeability across mouse ATII cell monolayers in ATII and THP1 coculture models. A , ATII cells were grown in 24-well transwell plates until the cellular monolayers reached confluence. THP-1 cells were added to the appropriate ATII monolayers, cocultured for 6 h, and exposed to hyperoxic conditions for 1 h. In some experiments, THP-1 cells were transfected with control shRNA or NALP3 shRNA and then added to a monolayer of ATII cells. Transepithelial albumin flux across each monolayer was determined by the addition of [ 125 I]-labeled human serum albumin to each upper compartment. The contents from the lower compartment were collected and counted in a Wizard γ-counter. * p

    Article Snippet: Transepithelial albumin flux across each monolayer was determined by the addition of 0.05 μCi [125 I]-labeled human serum albumin to each upper compartment and incubated for 1 h, after which the contents from the lower compartment were collected and counted in a Wizard γ-counter (Perkin-Elmer, Waltham, MA) as described previously ( ).

    Techniques: Permeability, Transfection, shRNA, Labeling

    Detection of tumor nodules with FDG-PET. (A) Gefitinib-treated and untreated A549 and HCC827 cells grown in 2D were incubated with 18 F-FDG for 60 minutes and FDG uptake was quantified using a gamma-counter. Data were corrected for background and decay and related to the initially added activity (n = 4, p = 0.02, Man Whitney U test). (B-I) Cross sections of lungs recellularized with tumor cells and incubated with 18 F-FDG were investigated by autoradiography (ARG). Regions of high radioactive intensity correlated with presence of tumor cells (arrows in B, C, D, E). Cells of both cell lines strongly expressed GLUT1 (F, G) while only a low amount of the tumor cells was proliferative as shown by Ki67 staining (H, I). Data are presented as arithmetic means ± SEM; *p

    Journal: PLoS ONE

    Article Title: Human Organotypic Lung Tumor Models: Suitable For Preclinical 18F-FDG PET-Imaging

    doi: 10.1371/journal.pone.0160282

    Figure Lengend Snippet: Detection of tumor nodules with FDG-PET. (A) Gefitinib-treated and untreated A549 and HCC827 cells grown in 2D were incubated with 18 F-FDG for 60 minutes and FDG uptake was quantified using a gamma-counter. Data were corrected for background and decay and related to the initially added activity (n = 4, p = 0.02, Man Whitney U test). (B-I) Cross sections of lungs recellularized with tumor cells and incubated with 18 F-FDG were investigated by autoradiography (ARG). Regions of high radioactive intensity correlated with presence of tumor cells (arrows in B, C, D, E). Cells of both cell lines strongly expressed GLUT1 (F, G) while only a low amount of the tumor cells was proliferative as shown by Ki67 staining (H, I). Data are presented as arithmetic means ± SEM; *p

    Article Snippet: 18 F-FDG uptake (2D cell cultures) 200,000 HCC827 or A549 cells were incubated with 1x106 counts per minute (cpm) of 18 F-FDG for 60 min. After incubation on ice to stop uptake and washing with PBS twice, remaining intracellular 18 F-FDG activity was quantified using a semi-automated gamma-counter (Wallac 1480-Wizard, Perkin Elmer, Germany).

    Techniques: Positron Emission Tomography, Incubation, Activity Assay, Autoradiography, Staining

    H2A monoubiquitination assay and CK2 kinase assay performed with γ- 32 P-ATP. a Coomassie blue staining of factors used. b Scheme for H2A monoubiquitination assay with E1 that was pre-charged with HA-ubiquitin (See Methods for details). c Immunoblotting of H2A monoubiquitination assay as describe in b with increasing amounts of CK2. d Radiograph of CK2 kinase assay reaction products. The assay was assembled with the factors indicated, each at the same amount used in the H2A monoubiquitination assay (Methods). After incubation at 37°C for 30 min, the assay was stopped by boiling in SDS loading buffer and resolved on SDS-PAGE. Besides CK2B, which was radio-labeled presumably due to autophosphorylation, phosphorylation of RING1B and PCGF5 was detected together with a species, indicated as *histone, dependent on the presence of nucleosomes. e H2A monoubquitination assay performed as in Fig. 3c , using increasing amounts of RING1B-PCGF5-AUTS2 containing either RING1B-S41A (S41 to alanine), or RING1B-S41D (S41 to aspartic acid), purified from sf9 cells.

    Journal: Nature

    Article Title: AUTS2 confers gene activation to Polycomb group proteins in the CNS

    doi: 10.1038/nature13921

    Figure Lengend Snippet: H2A monoubiquitination assay and CK2 kinase assay performed with γ- 32 P-ATP. a Coomassie blue staining of factors used. b Scheme for H2A monoubiquitination assay with E1 that was pre-charged with HA-ubiquitin (See Methods for details). c Immunoblotting of H2A monoubiquitination assay as describe in b with increasing amounts of CK2. d Radiograph of CK2 kinase assay reaction products. The assay was assembled with the factors indicated, each at the same amount used in the H2A monoubiquitination assay (Methods). After incubation at 37°C for 30 min, the assay was stopped by boiling in SDS loading buffer and resolved on SDS-PAGE. Besides CK2B, which was radio-labeled presumably due to autophosphorylation, phosphorylation of RING1B and PCGF5 was detected together with a species, indicated as *histone, dependent on the presence of nucleosomes. e H2A monoubquitination assay performed as in Fig. 3c , using increasing amounts of RING1B-PCGF5-AUTS2 containing either RING1B-S41A (S41 to alanine), or RING1B-S41D (S41 to aspartic acid), purified from sf9 cells.

    Article Snippet: CK2 (NEB, P6010) was then added at 300 nM along with 2 µCi γ-32 P-ATP (PerkinElmer, BLU502H500UC), followed by incubation at 37°C for 30 min.

    Techniques: Kinase Assay, Staining, Incubation, SDS Page, Labeling, Purification

    Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 P]ATP. The result of the reaction was run through an SDS-polyacrylamide

    Journal: The Journal of Biological Chemistry

    Article Title: Motor Domain Phosphorylation Modulates Kinesin-1 Transport *

    doi: 10.1074/jbc.M113.515510

    Figure Lengend Snippet: Estimation of phosphorylation efficiency using radiolabeling and SDS-PAGE. Phosphorylation was measured using radiolabeling. Kinesin was phosphorylated by JNK3 in the presence of [γ- 32 P]ATP. The result of the reaction was run through an SDS-polyacrylamide

    Article Snippet: This reaction was identical in composition to the nonradioactive mixture, but contained an additional 70 n m [γ-32 P]ATP (PerkinElmer Life Sciences, NEG502Z250UC).

    Techniques: Radioactivity, SDS Page

    Action of CK1δ within the purified nuclear PER complex (A) Protein kinase activity of nuclear PER complex. BN-PAGE autoradiogram showing control sample (WT) or affinity-purified nuclear PER complex (PER2-FH) after incubation with γ- 32 P-ATP (25°C, 1 h). . (C) CK1δ in purified PER complex phosphorylates PERs and CLOCK. SDS-PAGE autoradiogram as in (B). Leftmost lane, no inhibitor. Remaining lanes, decreasing concentrations (625, 125, 25, or 5 nM, represented by wedges) of PF-670462, a CK1δ/ε inhibitor (CK1δ/ε), or PF-4800567, a CK1ε-selective inhibitor (CK1ε). (D) Top and Middle, BN-APAGE immunoblots of mouse liver nuclear extracts (CT18) from wildtype (WT) or single mutants null for the circadian clock genes indicated at the top (KO) probed for proteins indicated at right. Bottom, SDS-PAGE SAP155 immunoblot loading control. (E) Top, SDS-PAGE autoradiogram of liver nuclear extracts (CT18) from genotypes indicated at top after immunoprecipitation with anti-CRY1 antibody and incubation with γ- 32 P-ATP (25°C, 10 min; linear range for wildtype). Bottom, SDS-PAGE immunoblot for CLOCK served as loading control for PER complexes. (F) Phosphor-imaging quantification of labeled CLOCK bands from (E) after normalization to CLOCK immunoblot signals from (E). Shown are mean ± SEM;  t -test (two-tailed). Dashed line, background.

    Journal: Molecular cell

    Article Title: Macromolecular assemblies of the mammalian circadian clock

    doi: 10.1016/j.molcel.2017.07.017

    Figure Lengend Snippet: Action of CK1δ within the purified nuclear PER complex (A) Protein kinase activity of nuclear PER complex. BN-PAGE autoradiogram showing control sample (WT) or affinity-purified nuclear PER complex (PER2-FH) after incubation with γ- 32 P-ATP (25°C, 1 h). . (C) CK1δ in purified PER complex phosphorylates PERs and CLOCK. SDS-PAGE autoradiogram as in (B). Leftmost lane, no inhibitor. Remaining lanes, decreasing concentrations (625, 125, 25, or 5 nM, represented by wedges) of PF-670462, a CK1δ/ε inhibitor (CK1δ/ε), or PF-4800567, a CK1ε-selective inhibitor (CK1ε). (D) Top and Middle, BN-APAGE immunoblots of mouse liver nuclear extracts (CT18) from wildtype (WT) or single mutants null for the circadian clock genes indicated at the top (KO) probed for proteins indicated at right. Bottom, SDS-PAGE SAP155 immunoblot loading control. (E) Top, SDS-PAGE autoradiogram of liver nuclear extracts (CT18) from genotypes indicated at top after immunoprecipitation with anti-CRY1 antibody and incubation with γ- 32 P-ATP (25°C, 10 min; linear range for wildtype). Bottom, SDS-PAGE immunoblot for CLOCK served as loading control for PER complexes. (F) Phosphor-imaging quantification of labeled CLOCK bands from (E) after normalization to CLOCK immunoblot signals from (E). Shown are mean ± SEM; t -test (two-tailed). Dashed line, background.

    Article Snippet: Briefly, in 100 μl of T4 Polynucleotide Kinase Reaction Buffer, 250 nM ssDNA EMSA Oligonucleotide was incubated with 5 μl of T4 polynucleotide kinase (NEB) and 18 μl of 3.3 μM 3000 Ci/mmol ATP, [γ-32 P] (Perkin-Elmer) for 1 h at 37°C Free ATP was removed using Illustra MicroSpin G-25 columns (GE). ssDNA was phenol-chloroform extracted.

    Techniques: Purification, Activity Assay, Polyacrylamide Gel Electrophoresis, Affinity Purification, Incubation, SDS Page, Western Blot, Immunoprecipitation, Imaging, Labeling, Two Tailed Test

    Reconstitution and 3D reconstruction of the PI3KC3-C1 complex. ( A ) Domain structures of the four subunits of PI3KC3-C1. The small unlabeled yellow box between VPS34 C2 and helical domains is the CHIL motif, described below. ( B ) Purification of PI3KC3-C1. Coomassie-stained SDS-PAGE gel of purified PI3KC3-C1. ( C ) Thin-layer chromatography of radiolabeled PI(3)P generated by PI3KC3-C1 from PI and [γ-32 P] ATP. ( D ) Reference-free class averages of PI3KC3-C1, each containing ∼200 particles. The first row represents particles that were selected for the 3D reconstruction. The lower two rows represent particles excluded from the 3D reconstruction, the arrowhead indicating the position of a density dislodged from the main part of the complex. All class averages shown are calculated from data acquired on untilted grids, except the top right class average which is from 45° tilt. ( E ) 3D reconstruction calculated from ∼39,000 particles in four orientations and displayed at a threshold determined from subsequent docking analyses. DOI: http://dx.doi.org/10.7554/eLife.05115.003

    Journal: eLife

    Article Title: Architecture and dynamics of the autophagic phosphatidylinositol 3-kinase complex

    doi: 10.7554/eLife.05115

    Figure Lengend Snippet: Reconstitution and 3D reconstruction of the PI3KC3-C1 complex. ( A ) Domain structures of the four subunits of PI3KC3-C1. The small unlabeled yellow box between VPS34 C2 and helical domains is the CHIL motif, described below. ( B ) Purification of PI3KC3-C1. Coomassie-stained SDS-PAGE gel of purified PI3KC3-C1. ( C ) Thin-layer chromatography of radiolabeled PI(3)P generated by PI3KC3-C1 from PI and [γ-32 P] ATP. ( D ) Reference-free class averages of PI3KC3-C1, each containing ∼200 particles. The first row represents particles that were selected for the 3D reconstruction. The lower two rows represent particles excluded from the 3D reconstruction, the arrowhead indicating the position of a density dislodged from the main part of the complex. All class averages shown are calculated from data acquired on untilted grids, except the top right class average which is from 45° tilt. ( E ) 3D reconstruction calculated from ∼39,000 particles in four orientations and displayed at a threshold determined from subsequent docking analyses. DOI: http://dx.doi.org/10.7554/eLife.05115.003

    Article Snippet: The reaction was started by adding 6 µl cold ATP (0.5 mM in reaction buffer) and 1 µl ATP [γ-32P] (10 µCi, PerkinElmer).

    Techniques: Purification, Staining, SDS Page, Thin Layer Chromatography, Generated

    PI5P4Kβ has ability to sense GTP concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM γ- 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .

    Journal: Molecular cell

    Article Title: The lipid kinase PI5P4Kβ is an intracellular GTP sensor for metabolism and tumorigenesis

    doi: 10.1016/j.molcel.2015.12.011

    Figure Lengend Snippet: PI5P4Kβ has ability to sense GTP concentration (A) PI5P4Kβ utilizes GTP as a preferred phosphodonor to produce PI(4,5)P 2 from PI(5)P. The bacterially produced recombinant PI5P4Kβ was incubated with PI(5)P in the presence of 0.2 µM γ- 32 P radiolabeled GTP or ATP. The phospholipids were extracted and analyzed by thin layer chromatography and autoradiography. (B) PI5P4Kβ activity changes within the physiological variations of GTP concentration. PI5P4Kβ kinase activity was assessed with the indicated levels of γ- 32 P radiolabeled GTP or ATP and 20 µM of PI(5)P. (C) PI5P4Kβ retains its preference to GTP at any PI(5)P concentration. (D) PI5P4Kβ retains GTP responsiveness in the presence of physiological amount of ATP. PI5P4Kβ kinase activity with the indicated amount of γ- 32 P radiolabeled GTP was assessed in the presence of 1 mM and 2 mM of cold (non-radiolabeled) ATP. (E) PI5P4Kβ efficiently utilizes GTP in the presence of physiological amount of ATP. GTP- and ATP-dependent kinase activities of PI5P4Kβ were assessed with mixtures of cold and γ- 32 P radiolabeled ATP and GTP as shown in the left panel. 0.2 µg PI5P4Kβ and 1.0 µg PI(5)P was used for each reaction and only radiolabeled PI(4,5)P 2 was quantified (right panel). Data are displayed as means ± SD, n=3. .

    Article Snippet: The in vitro kinase assay was carried out using γ-32 P radiolabeled ATP or GTP (Perkin Elmer), synthetic PI(5)P diC16 (Echelon Biosciences), and recombinant PI5P4Ks.

    Techniques: Concentration Assay, Produced, Recombinant, Incubation, Thin Layer Chromatography, Autoradiography, Activity Assay

    GTP-sensing activity of PI5P4Kβ regulates metabolism and PI(5)P level (A) The comparable expression levels of WT-PI5P4Kβ or PI5P4Kβ F205L in the isogenic PI5P4Kβ null MEFs. (B) Cellular GTP and ATP concentrations of WT-PI5P4Kβ and PI5P4Kβ F205L cells with or without MPA treatment for 4hr. The average and deviation of two independent samples per group is presented. The data are representative of two separate experiments. (C) Heat map of the metabolic responses to the GTP concentration changes. Each metabolite is color coded by the changes in amount by the MPA treatment. (D) Cellular PI(5)P levels of WT-PI5P4Kβ and PI5P4Kβ F205L cells with or without MPA treatment for 4 hr. The average and deviation of two independent samples per group is presented. The data are representative of two separate experiments. .

    Journal: Molecular cell

    Article Title: The lipid kinase PI5P4Kβ is an intracellular GTP sensor for metabolism and tumorigenesis

    doi: 10.1016/j.molcel.2015.12.011

    Figure Lengend Snippet: GTP-sensing activity of PI5P4Kβ regulates metabolism and PI(5)P level (A) The comparable expression levels of WT-PI5P4Kβ or PI5P4Kβ F205L in the isogenic PI5P4Kβ null MEFs. (B) Cellular GTP and ATP concentrations of WT-PI5P4Kβ and PI5P4Kβ F205L cells with or without MPA treatment for 4hr. The average and deviation of two independent samples per group is presented. The data are representative of two separate experiments. (C) Heat map of the metabolic responses to the GTP concentration changes. Each metabolite is color coded by the changes in amount by the MPA treatment. (D) Cellular PI(5)P levels of WT-PI5P4Kβ and PI5P4Kβ F205L cells with or without MPA treatment for 4 hr. The average and deviation of two independent samples per group is presented. The data are representative of two separate experiments. .

    Article Snippet: The in vitro kinase assay was carried out using γ-32 P radiolabeled ATP or GTP (Perkin Elmer), synthetic PI(5)P diC16 (Echelon Biosciences), and recombinant PI5P4Ks.

    Techniques: Activity Assay, Expressing, Concentration Assay

    PI5P4Kβ is a GTP-binding phosphatidylinositol kinase (A) A list for kinases that were identified in a proteome screening with GTP-conjugated agarose beads. (B) Enzymatic functions of PI5P4K/Type II PIPK and PI4P5K/Type I PIPK. (C) Cellular PI5P4Kβ binds to GTP-conjugated agarose beads more than ATP- and m7GTP-conjugated agarose beads. After incubated with HEK 293T cell lysates, PI5P4Kβ as well as control proteins that bound to the indicated control or nucleotide conjugated beads were immunoblotted. . (E) PI5P4Kβ possesses superior GTP hydrolysis activity in vitro .

    Journal: Molecular cell

    Article Title: The lipid kinase PI5P4Kβ is an intracellular GTP sensor for metabolism and tumorigenesis

    doi: 10.1016/j.molcel.2015.12.011

    Figure Lengend Snippet: PI5P4Kβ is a GTP-binding phosphatidylinositol kinase (A) A list for kinases that were identified in a proteome screening with GTP-conjugated agarose beads. (B) Enzymatic functions of PI5P4K/Type II PIPK and PI4P5K/Type I PIPK. (C) Cellular PI5P4Kβ binds to GTP-conjugated agarose beads more than ATP- and m7GTP-conjugated agarose beads. After incubated with HEK 293T cell lysates, PI5P4Kβ as well as control proteins that bound to the indicated control or nucleotide conjugated beads were immunoblotted. . (E) PI5P4Kβ possesses superior GTP hydrolysis activity in vitro .

    Article Snippet: The in vitro kinase assay was carried out using γ-32 P radiolabeled ATP or GTP (Perkin Elmer), synthetic PI(5)P diC16 (Echelon Biosciences), and recombinant PI5P4Ks.

    Techniques: Binding Assay, Incubation, Activity Assay, In Vitro

    Representative gamma images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.

    Journal: Molecules

    Article Title: Design, Synthesis and Evaluation of Dual-Modality Glyco-Nanoparticles for Tumor Imaging

    doi: 10.3390/molecules18066425

    Figure Lengend Snippet: Representative gamma images of A 431 bearing nude mice at various times (left: 111 In-DTPA-NIRF-CCPM; right: 111 In-DG-CCPM). Arrows indicate the presence of tumors.

    Article Snippet: Radioactivity was counted using an automated gamma scintillation counter (Perkin Elmer, 1470-002, Boston, MA, USA).

    Techniques: Mouse Assay

    Immunomodulatory effect of endogenous IL-18 and IL-12 on intrapulmonary IFN-γ during replicative L. pneumophila lung infection. A/J mice were administered anti-IL-18R MAb, anti-IL-12 MAb, or both MAbs prior to intratracheal inoculation with L. pneumophila . At specific time points p.i., mice were euthanatized, and lungs were excised, homogenized, and filtered. Intrapulmonary levels of IFN-γ were quantified in filtered lung homogenates by ELISA. Mice were treated with control antibody (IgG2a) (solid bars), anti-IL-12 MAb (shaded bars), anti-IL-18RmAB (hatched bars), anti-IL-12 MAb and anti-IL-18R MAb (horizontally striped bars). Results represent means ± standard errors of the means for five animals per treatment group. ∗, P

    Journal: Infection and Immunity

    Article Title: Immunomodulatory Role of Endogenous Interleukin-18 in Gamma Interferon-Mediated Resolution of Replicative Legionella pneumophila Lung Infection

    doi:

    Figure Lengend Snippet: Immunomodulatory effect of endogenous IL-18 and IL-12 on intrapulmonary IFN-γ during replicative L. pneumophila lung infection. A/J mice were administered anti-IL-18R MAb, anti-IL-12 MAb, or both MAbs prior to intratracheal inoculation with L. pneumophila . At specific time points p.i., mice were euthanatized, and lungs were excised, homogenized, and filtered. Intrapulmonary levels of IFN-γ were quantified in filtered lung homogenates by ELISA. Mice were treated with control antibody (IgG2a) (solid bars), anti-IL-12 MAb (shaded bars), anti-IL-18RmAB (hatched bars), anti-IL-12 MAb and anti-IL-18R MAb (horizontally striped bars). Results represent means ± standard errors of the means for five animals per treatment group. ∗, P

    Article Snippet: Primers for IL-12 and IFN-γ were obtained from Perkin-Elmer as predeveloped assay reagents (PDARs).

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Kinetics of intrapulmonary IL-12 and IFN-γ induction during replicative L. pneumophila lung infection. A/J mice were inoculated with virulent L. pneumophila (10 6 CFU/mouse). At specific time points p.i., the mice were euthanatized, and the lungs were excised. Total RNA was extracted, or the lungs were lavaged for collection of BALF. Transcript levels were quantified by real-time RT-PCR (IL-12p40 [a], IL-12p35 [b], and IFN-γ [c], while protein levels were quantified in BALF by cytokine-specific ELISA (IL-12p70 [d] and IFN-γ [e]). For mRNA quantification, PCR amplification of the housekeeping gene ubiquitin was performed for each sample to control for sample loading and to facilitate normalization between samples. The ubiquitin-normalized data was expressed as the fold induction of gene expression of L. pneumophila -infected mice compared to uninfected mice. Results represent fold induction of gene expression from six to eight mice per time point. For quantification of cytokines in BALF, results represent the means ± standard errors of the means of three to five animals per time point. ∗, P

    Journal: Infection and Immunity

    Article Title: Immunomodulatory Role of Endogenous Interleukin-18 in Gamma Interferon-Mediated Resolution of Replicative Legionella pneumophila Lung Infection

    doi:

    Figure Lengend Snippet: Kinetics of intrapulmonary IL-12 and IFN-γ induction during replicative L. pneumophila lung infection. A/J mice were inoculated with virulent L. pneumophila (10 6 CFU/mouse). At specific time points p.i., the mice were euthanatized, and the lungs were excised. Total RNA was extracted, or the lungs were lavaged for collection of BALF. Transcript levels were quantified by real-time RT-PCR (IL-12p40 [a], IL-12p35 [b], and IFN-γ [c], while protein levels were quantified in BALF by cytokine-specific ELISA (IL-12p70 [d] and IFN-γ [e]). For mRNA quantification, PCR amplification of the housekeeping gene ubiquitin was performed for each sample to control for sample loading and to facilitate normalization between samples. The ubiquitin-normalized data was expressed as the fold induction of gene expression of L. pneumophila -infected mice compared to uninfected mice. Results represent fold induction of gene expression from six to eight mice per time point. For quantification of cytokines in BALF, results represent the means ± standard errors of the means of three to five animals per time point. ∗, P

    Article Snippet: Primers for IL-12 and IFN-γ were obtained from Perkin-Elmer as predeveloped assay reagents (PDARs).

    Techniques: Infection, Mouse Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Amplification, Expressing

    Cdk1 is important for ESC pluripotency and preferentially interacts with Oct4 at the G2/M phase. ( A ) Protein levels and mRNA levels of Cdk1-knockdown mESCs ( n = 3). ( B and C ) AP staining of Cdk1-knockdown mESCs. After staining to reveal AP activity, the colonies were scored and the percentages of undifferentiated, partially differentiated, and fully differentiated colonies were calculated. AP staining experiments were repeated three times ( n = 3). Scale bar represents 500 μm. ( D ) Immunostaining of Cdk1-knockdown mESCs. Cdk1 was stained with anti-Cdk1 (green) and DNA was stained DAPI (blue). Scale bar represents 100 μm. ( E ) Fluorescence images of Cdk1-knockdown mESCs. Nanog and Klf4 were stained with anti-Nanog (green) and anti-Klf4 (green), respectively. DNA was stained with DAPI (blue). Scale bar represents 100 μm. ( F ) Flag-Oct4 and HA-Cdk1 were cotransfected into HEK293T cells. Cell lysates were immunoprecipitated with anti-Flag antibody and probed with anti-HA antibody (left). Endogenous Cdk1 was immunoprecipitated from E14 mESCs with anti-Oct4 antibody and immunoblotted with anti-Cdk1 antibody (right). ( G ) Changes in interaction of Cdk1 with Oct4 during cell cycle progression. Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole (100 ng/ml) for 8 h. At the indicated time after release into fresh media, cell cycle progression was determined by FACS analysis (right). Cell lysates were pulled down with anti-Flag beads. Bound proteins were eluted and then immunoblotted with the indicated antibodies. (A, asynchronous state; N, nocodazole treatment). ( H ) Changes in interaction of Oct4 with Cdk1 depending on Cyclin B knockdown. After Cyclin B knockdown, Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole for 8 h. Flag-Oct4 was immunoprecipitated by incubating lysates with anti-Flag beads. Following Flag elution, bound proteins were immunoblotted with the indicated antibodies. ( I ) Colocalization of Cdk1 and p-Oct4(S229) in mitosis-arrested mESCs was analyzed by immunostaining. mESCs were treated with nocodazole (100 ng/ml) for 8 h. Cdk1 was stained with anti-Cdk1 (green), p-Oct4(S229) was stained with anti-p-Oct4(S229) (red), and DNA was stained with DAPI (blue). Scale bar represents 5 μm. ( J ) Radioactive in vitro kinase assay using recombinant Cdk1 to phosphorylate GST-Oct4 wild type (WT) and S228A/S229A mutant. Autoradiogram showing incorporation of γ- 32 P ATP (left). For the IP kinase assay, cell lysates of nocodazole-treated E14 mESCs were immunoprecipitated with anti-CycB1 and Aurkb and subjected to an in vitro kinase assay using (His) 6 -PP1α and GST-Oct4 as a substrate and followed by western blotting (right).

    Journal: Nucleic Acids Research

    Article Title: Cyclin-dependent kinase 1 activity coordinates the chromatin associated state of Oct4 during cell cycle in embryonic stem cells

    doi: 10.1093/nar/gky371

    Figure Lengend Snippet: Cdk1 is important for ESC pluripotency and preferentially interacts with Oct4 at the G2/M phase. ( A ) Protein levels and mRNA levels of Cdk1-knockdown mESCs ( n = 3). ( B and C ) AP staining of Cdk1-knockdown mESCs. After staining to reveal AP activity, the colonies were scored and the percentages of undifferentiated, partially differentiated, and fully differentiated colonies were calculated. AP staining experiments were repeated three times ( n = 3). Scale bar represents 500 μm. ( D ) Immunostaining of Cdk1-knockdown mESCs. Cdk1 was stained with anti-Cdk1 (green) and DNA was stained DAPI (blue). Scale bar represents 100 μm. ( E ) Fluorescence images of Cdk1-knockdown mESCs. Nanog and Klf4 were stained with anti-Nanog (green) and anti-Klf4 (green), respectively. DNA was stained with DAPI (blue). Scale bar represents 100 μm. ( F ) Flag-Oct4 and HA-Cdk1 were cotransfected into HEK293T cells. Cell lysates were immunoprecipitated with anti-Flag antibody and probed with anti-HA antibody (left). Endogenous Cdk1 was immunoprecipitated from E14 mESCs with anti-Oct4 antibody and immunoblotted with anti-Cdk1 antibody (right). ( G ) Changes in interaction of Cdk1 with Oct4 during cell cycle progression. Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole (100 ng/ml) for 8 h. At the indicated time after release into fresh media, cell cycle progression was determined by FACS analysis (right). Cell lysates were pulled down with anti-Flag beads. Bound proteins were eluted and then immunoblotted with the indicated antibodies. (A, asynchronous state; N, nocodazole treatment). ( H ) Changes in interaction of Oct4 with Cdk1 depending on Cyclin B knockdown. After Cyclin B knockdown, Flag-Oct4-expressing ZHBTc4 mESCs were treated with nocodazole for 8 h. Flag-Oct4 was immunoprecipitated by incubating lysates with anti-Flag beads. Following Flag elution, bound proteins were immunoblotted with the indicated antibodies. ( I ) Colocalization of Cdk1 and p-Oct4(S229) in mitosis-arrested mESCs was analyzed by immunostaining. mESCs were treated with nocodazole (100 ng/ml) for 8 h. Cdk1 was stained with anti-Cdk1 (green), p-Oct4(S229) was stained with anti-p-Oct4(S229) (red), and DNA was stained with DAPI (blue). Scale bar represents 5 μm. ( J ) Radioactive in vitro kinase assay using recombinant Cdk1 to phosphorylate GST-Oct4 wild type (WT) and S228A/S229A mutant. Autoradiogram showing incorporation of γ- 32 P ATP (left). For the IP kinase assay, cell lysates of nocodazole-treated E14 mESCs were immunoprecipitated with anti-CycB1 and Aurkb and subjected to an in vitro kinase assay using (His) 6 -PP1α and GST-Oct4 as a substrate and followed by western blotting (right).

    Article Snippet: For radioactive in vitro kinase assay, (His)6 -PP1 and GST-Oct4 were incubated with Cdk1/CyclinB1 in kinase buffer (60 mM HEPES–NaOH pH 7.5, 3 mM MgCl2 , 3 mM MnCl2 , 3 mM Na-orthovanadate, 1.2 mM DTT, 0.25 mM ATP) with 0.1 mM γ-32 P-ATP (NEG002A250UC, purchased from PerkinElmer, Waltham, MA, USA) for 30 min at 30°C.

    Techniques: Staining, Activity Assay, Immunostaining, Fluorescence, Immunoprecipitation, Expressing, FACS, In Vitro, Kinase Assay, Recombinant, Mutagenesis, IP-Kinase Assay, Western Blot