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0.000005 - 0.000076
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depending on presence of divalent cations
0.0051
Murine autotaxin gamma, p-nitrophenyl phenylphosphonate as a substrate, Vmax 1.8 nmol p-nitrophenyl/min
0.00847
human autotaxin alpha, p-nitrophenyl phenylphosphonate as a substrate, Vmax 0.67 nmol p-nitrophenyl/min
0.0163
Murine autotaxin alpha, p-nitrophenyl phenylphosphonate as a substrate, Vmax 0.35 nanomol p-nitrophenyl/min
0.036
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using isotopic assay
0.048
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colorimetric assay, 6 h incubation, 37°C
0.0919
Murine autotaxin beta, para-nitrophenyl phenylphosphonate as a substrate, Vmax 1.9 nmol para-nitrophenyl/min
0.0997
Human autotaxin gamma, p-nitrophenyl phenylphosphonate as a substrate, Vmax 1.6 nmol p-nitrophenyl/min
0.135
Human autotaxin beta, p-nitrophenyl phenylphosphonate as a substrate, Vmax 1.9 nmol p-nitrophenyl/min
0.14
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purified enzyme, HiTrap DEAE FF
additional information
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Spectrophotometric assay of LysoPLD activity. Determination of lysoPLD activity by measuring the amount of choline released from exogenously added LPC. Fluorometric assay for LysoPLD activity. Determination of LysoPLD activity using a fluorogenic substrate, [5-[[(23S)-33-(4-[(E)-[4-(dimethylamino)phenyl]diazenyl]phenyl)-20,23-dihydroxy-20-oxido-15,26,33-trioxo-3,6,9,12,19,21,25-heptaoxa-16,32-diaza-20-phosphatritriacont-1-yl]carbamoyl]-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl]acetic acid (FS-3)
additional information
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lysoPLD activity of diluted or undiluted egg white measured based on enzyme-linked fluorometric measurement of choline produced together with lysophosphatidic acid from exogenous lysophosphatidylcholine. LysoPLD activity measured by determining lysophosphatidic acid and choline by MALDI-TOF-MS, mass spectrometric and enzyme-linked fluorometric analysis. Quantitative analysis of lysophosphatidic acid by MALDI-TOF-MS using a phosphate-capture molecule. Production of lysophosphatidic acid during incubation of hen egg white, changes in the levels of fatty acyl groups
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autotaxin/lysoPLD activity is determined at the indicated substrate concentrations using conditioned medium from V5-autotaxin expressing cells as the source of enzyme and radiolabeled lysophosphatidylcholine as substrate
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biochemical and cell biological activity of hATX S48 protein measured
additional information
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biochemical and cell biological activity of hATX S48 protein measured
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lysoPLD activity is assessed, based on the amount of choline released with lysophosphatidylcholine as the substrate, absorption spectrometry. Amounts of the substrate lysophosphatidylcholine and the product lysophosphatidic acid are also checked simultaneously. Changes in the lysoPLD activity and lysophosphatidylcholine and lysophosphatidic acid concentrations in incubated serum. Concentrations of lysophosphatidylcholine and lysophosphatidic acid increase upon incubation, the levels increase significantly as compared with those in the controls (no incubation) after 60-min incubations. Dramatic increase in the lysophosphatidic acid concentration is observed, and the lysophosphatidic acid level after 180-min incubation is about 15times higher than in the control
additional information
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phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl phenylphosphonate substrate
additional information
phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl phenylphosphonate substrate
additional information
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approx. 0 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
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approx. 0 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 0 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
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approx. 10 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 100 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 1000 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 104 micromol/ml/min, serum from a patient with severe pre-eclampsia, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 1100 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 118 micromol/ml/min, serum from a patient with preterm labor (30.1 +/- 0.8 gestational weeks; n is 21), diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
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approx. 139 micromol/ml/min, serum from a patient with preterm labor (30.1 +/- 0.8 gestational weeks; n is 21), diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 18:2-lysophosphatidylcholine, radiolabeled substrate, 37°C
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approx. 146 micromol/ml/min, serum from a patient with severe pre-eclampsia, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 18:2-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 15 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 15 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
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approx. 150 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
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approx. 153 micromol/ml/min, serum from a woman with normal pregnancy (35.8 +/- 0.7 gestational weeks; n is 25), diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 18:2-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 180 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 200 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 300 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 350 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 400 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 450 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 450 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 450 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
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approx. 49 micromol/ml/min, serum from a patient with moderate pre-eclampsia, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 18:2-lysophosphatidylcholine, radiolabeled substrate, 37°C
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approx. 5 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 500 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 500 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 60 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 65 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 65 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 65 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 1fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 700 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 100fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 8 mmol/ml/min, serum from a patient with preterm labor, detection of dilution dependency of choline-producing activity of sera, diluted 30fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 8 mmol/ml/min, serum from a woman with normal pregnancy, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
additional information
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approx. 80 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 3.3fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 90 micromol/ml/min, serum from a woman with normal pregnancy (35.8 +/- 0.7 gestational weeks; n is 25), diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx. 92 micromol/ml/min, serum from a patient with moderate pre-eclampsia, diluted 3.3fold (serum + saline), presence of 0.15 mM exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate, 37°C
additional information
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approx.50 mmol/ml/min, serum from a patient with severe pre-eclampsia, detection of dilution dependency of choline-producing activity of sera, diluted 10fold (serum + saline), absence of exogenous 16:0-lysophosphatidylcholine, radiolabeled substrate , 37°C
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overview of methods for determination of autotaxin/lysoPLD activity. Using radiolabeled substrates and fluorogenic substrates for measuring autotaxin/lysoPLD activity
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lysophospholipase D activity is measured by conversion of radiolabeled lysophosphatidylcholine into radiolabeled lysophosphatidic acid
additional information
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phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl phenylphosphonate substrate
additional information
phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl phenylphosphonate substrate
additional information
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phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl-phenylphosphonate substrate
additional information
phosphodiesterase activity of recombinant autotaxin is measured by using p-nitrophenyl-phenylphosphonate substrate
additional information
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phosphodiesterase activity of recombinant autotaxin is measured by using para-nitrophenyl phenylphosphonate substrate
additional information
phosphodiesterase activity of recombinant autotaxin is measured by using para-nitrophenyl phenylphosphonate substrate
additional information
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To deplete ATX from mouse serum, mouse serum is incubated with the 5E5-Sepharose 4B for 2 h at 4°C, and the resulting supernatant is used for measuring lysoPLD activity and lysophosphatidic acid production. Lysophospholipase D activity of plasma and amniotic fluids isolated from atx+/+ and atx+/-adult mice or atx+/+ and atx+/-embryos at different stages of development measured with immunohistochemistry. LysoPLD activity is determined by liberation of choline from lysophosphatidylcholine using myristyl-lysophosphatidylcholine as a substrate
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approx. 400 micromol/min/ml, mice are treated with control rat IgG, plasma samples (10 microl) incubate with 4 mmol/l lysophosphatidylcholine, activity determined by enzymatic photometric method using choline oxidase at 560 nm, 37°C
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approx. 42 micromol/min/ml, mice are treated with anti-ATX mAb, plasma samples (10 microl) incubate with 4 mmol/l lysophosphatidylcholine, activity determined by enzymatic photometric method using choline oxidase at 560 nm, 37°C
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nuclear fraction has relatively high levels of lysoPLD activity. Both lysoPLD activity and highest band density found in microsomal fraction