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Gly pro aminoluciferin

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Gly-Pro-aminoluciferin is a substrate for the measurement of prolyl oligopeptidase (POP) enzyme activity in biological samples. It is a chemiluminescent compound that generates light upon cleavage by POP, allowing for the quantitative detection of POP levels.

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Lab products found in correlation

Xenogen ivis system, by PerkinElmer (2 mentions) Dppiv glo protease assay, by Promega (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Glomax multi detection system, by Promega (1 mentions)

3 protocols using gly pro aminoluciferin

1

Evaluating DPP4 Activity in Mice

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Example 4

For evaluation of DPP4 enzymatic activity, mice were bled at the time points indicated and plasma samples were collected after centrifugation of blood. For measurements of DPP4 activity in tumor homogenates, tumors growing in mice were dissected, weighted and homogenized in PBS supplemented with protease inhibitor cocktail (Roche). Soluble extracts were collected after centrifugation of tumor homogenates. DPP4 activity in peritoneal cavity was measured by collecting peritoneal washes in 1.5 ml of PBS. DPP4 activity was measured using the DPPIV-Glo™ Protease assay (Promega). To evaluate DPP4 activity in vivo, FVB-Tg(CAG-luc) mice were fed with control and sitagliptin chow 24 hours prior to intraperitoneal injection of 10 mM Gly-Pro-aminoluciferin (Promega). Bioluminescence images were acquired with a XENOGEN (IVIS system, Perkin Elmer), 5 min after injection.

US11000521B2. Dipeptidylpeptidase 4 inhibition enhances lymphocyte trafficking, improving both naturally occurring tumor immunity and immunotherapy (2021-05-11). INSTITUT PASTEUR [FR], INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE (INSERM) [FR]. Inventors: Rosa Barreira Da Silva [US], Matthew Albert [FR].
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2

DPP4 Activity Measurement in AML Exosomes

Cited 1 time
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Aliquots of exosomes (10 μg protein) isolated from AML patients’ plasma or leukemia cell lines were suspended in 50 μL PBS and plated in wells of a 96-well flat-bottom microtiter plate. The assay was carried out by combining the exosomes with 50 μL of the DPP4 substrate, Gly-Pro-aminoluciferin, and the buffer optimized for this assay (Promega, Madison, WI, USA). Plates were incubated at 37 °C for 60 min, and surface DPP4 enzyme activity was measured as luminescence signals using the Promega GloMax multidetection system. Diprotin A, a DPP4 inhibitor, was used for blocking DPP4 activity [24 (link)]. Titration experiments were performed using different doses of Diprotin A (0.1–0.6 mmol) to identify the optimal inhibitory dose of Diprotin A for the blocking experiments. Aliquots of exosomes (10 μg protein) were co-incubated with Diprotin A (0.2 mmol) for 30 min prior to the addition of DPP4.
Namburi S., Broxmeyer H.E., Hong C.S., Whiteside T.L, & Boyiadzis M. (2020). DPP4+ exosomes in AML patients’ plasma suppress proliferation of hematopoietic progenitor cells. Leukemia, 35(7), 1925-1932.
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3

Measuring DPP4 Activity in Mice

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Example 4

For evaluation of DPP4 enzymatic activity, mice were bled at the time points indicated and plasma samples were collected after centrifugation of blood. For measurements of DPP4 activity in tumor homogenates, tumors growing in mice were dissected, weighted and homogenized in PBS supplemented with protease inhibitor cocktail (Roche). Soluble extracts were collected after centrifugation of tumor homogenates. DPP4 activity in peritoneal cavity was measured by collecting peritoneal washes in 1.5 ml of PBS. DPP4 activity was measured using the DPPIV-Glo™ Protease assay (Promega). To evaluate DPP4 activity in vivo, FVB-Tg(CAG-luc) mice were fed with control and sitagliptin chow 24 hours prior to intraperitoneal injection of 10 mM Gly-Pro-aminoluciferin (Promega). Bioluminescence images were acquired with a XENOGEN (IVIS system, Perkin Elmer), 5 min after injection.

US11229645B2. Dipeptidylpeptidase 4 inhibition enhances lymphocyte trafficking, improving both naturally occurring tumor immunity and immunotherapy (2022-01-25). INSTITUT PASTEUR [FR], INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE (INSERM) [FR]. Inventors: Rosa Barreira Da Silva [US], Matthew Albert [FR].
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