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Mcpt1

Manufactured by Thermo Fisher Scientific
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Mcpt1 is a laboratory instrument designed for the detection and quantification of mast cell proteases. It utilizes immunoassay technology to measure the levels of specific mast cell proteases in biological samples. The core function of Mcpt1 is to provide accurate and reliable data on mast cell protease levels, which can be useful in research and clinical applications.

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Lab products found in correlation

Mcpt4, by Aviva Systems Biology (5 mentions) Ova specific igg1, by Alpha Diagnostic (4 mentions) Bio plex pro mouse cytokine il 33 set, by Bio-Rad (2 mentions) Microplate reader, by BMG Labtech (2 mentions) Histamine, by Enzo Life Sciences (2 mentions) Microplate reader, by Molecular Devices (2 mentions) Procartaplex immunoassay, by Thermo Fisher Scientific (1 mentions) Igg2a, by Thermo Fisher Scientific (1 mentions) Cdna archive reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Ajap1, by Thermo Fisher Scientific (1 mentions) Kcnmb4, by Thermo Fisher Scientific (1 mentions) Msr 1, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Bms618 3, by Thermo Fisher Scientific (1 mentions) Celltiter glo assay, by Promega (1 mentions) Streptavidin horseradish peroxidase hrp, by BD (1 mentions) Protease inhibitor cocktail, by Fujifilm (1 mentions) Tmb substrate solution, by BD (1 mentions) Hmgb 1, by Promega (1 mentions)

14 protocols using mcpt1

1

Quantifying Serum Immunoglobulin Levels

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Serum concentrations of IgE (Biolegend, #432404), IgM (Thermo Fisher Scientific, #88-50470-88), IgA (Thermo Fisher Scientific, #88-50450-86), IgG1 (Thermo Fisher Scientific, #88-50410-86) IgG2a (Thermo Fisher Scientific, #88-50420-22), and MCPT-1 (Thermo Fisher Scientific, #88-7503-22) were measured by ELISA kits. ProcartaPlex Immunoassays (Thermo Fisher Scientific, #EPX070-20815-901) was used for measuring serum Igs in Supplementary Fig. 2A according to the manufacturer’s instructions.
Kwon D.I., Park E.S., Kim M., Choi Y.H., Lee M.S., Joo S.H., Kang Y.W., Lee M., Jo S.B., Lee S.W., Kim J.K, & Lee Y.J. (2022). Homeostatic serum IgE is secreted by plasma cells in the thymus and enhances mast cell survival. Nature Communications, 13, 1418.
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2

Comparative ADSC Gene Expression

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Gene expression in young, aged ADSCs was analyzed by quantitative real-time RT-PCR in technical triplicates using SYBR Green Premix Ex Taq II (Tli RNaseH Plus). Brie y, total RNA was extracted using Qiagen RNeasy Mini kit (Qiagen, Valencia, CA), following the manufacturer' s protocols. cDNA was synthesized using the cDNA archive reverse transcription kit (Life Technologies). Primers for Lcn2, Msr1, Tyrobp, Nfasc, Ccl12, Lef1, Mcpt1, Dthd1, Ajap1, Kcnmb4 were purchased from ThermoFisher. Actin was used as the reference gene for normalization. The cycle threshold (Ct) method of relative quanti cation of gene expression was used for these PCRs (ΔΔCt). The speci c primers are listed in Supporting Information Table S1.
Li K., Shi G., Lei X., Huang Y., Li X., Bai L., & Qin C. (2021). The CCL7-CCL2-CCR2 Axis Regulates Age-Related Alterations in ADSCs .
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3

Quantifying Allergy-related Biomarkers in Mice

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Serum samples from blood drawn after cardiac puncture were analyzed using ELISA kits for OVA-specific IgE (MD Bioproducts, Oakdale, MN, USA), MCPt-1 (Invitrogen, Carlsbad, CA, USA), and OVA-specific IgG1 (Alpha Diagnostic International, San Antonio, TX, USA). Total IgE (Bioscience, San Diego, CA, USA) was performed for steady state analysis. For intestinal histological analyses, duodenal tissue was fixed in 10% formalin and processed by standard histological procedures as previously described [36 (link)]. At least eight random images were taken from at least three random sections per mouse. Quantification of stained cells per square millimeter was performed by morphometric analysis using Image Processing Software (ImagePro, Media Cybernetics, MD, USA).
Sharma S., Tomar S., Dharne M., Ganesan V., Smith A., Yang Y., Waggoner L., Wang Y.H, & Hogan S.P. (2019). Deletion of ΔdblGata motif leads to increased predisposition and severity of IgE-mediated food-induced anaphylaxis response. PLoS ONE, 14(8), e0219375.
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4

Quantification of Cytokines and IgE via ELISA

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IL-9 (Biolgend), MCPT-1, and IgE (Invitrogen) enzyme-linked immunosorbent assays (ELISAs) were performed according to the manufacturer’s instruction. Briefly, 96-well plates were coated with coating antibody overnight at 4 °C. After washing three times with the wash buffer, 300 μl ELISA buffer was added to the plate and incubate at room temperature for 2 h. After washing three times with washing buffer, 100 μl samples were added to the plate and incubated at room temperature for 2 h. After washing for three times, 100 μl diluted detection antibody was added to the plate and incubated at room temperature for 1 h. After washing the plate three times, 100 μl of diluted Avidin-horseradish peroxidase solution was added to the plate and incubated at room temperature for 30 min in the dark. After washing the plate for three times, 100 μl substrate was added to the plate. Plates were read at absorbance 450 nm and 570 nm. Details of antibodies are listed in Supplementary Table 9.
Fu Y., Wang J., Panangipalli G., Ulrich B.J., Koh B., Xu C., Kharwadkar R., Chu X., Wang Y., Gao H., Wu W., Sun J., Tepper R.S., Zhou B., Janga S.C., Yang K, & Kaplan M.H. (2020). STAT5 promotes accessibility and is required for BATF-mediated plasticity at the Il9 locus. Nature Communications, 11, 4882.
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5

Quantifying Intestinal Mast Cells and Goblet Cells

Cited 2 times
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To measure intestinal mast cell number and levels of goblet cell hyperplasia, duodenal tissue was fixed in 10% formalin and processed by standard histological techniques. 5–8–μm tissue sections were stained with Leder stain for chloroacetate esterase (CAE) activity in intestinal mast cells or periodic acid-Schiff (PAS) for mucins in goblet cells. Stained cells were quantified as previously described3 (link). To measure secreted mediators, serum samples were analyzed using ELISA kits of OVA-specific IgE (MD Bioproducts), MCPt-1 (eBioscience), and OVA-specific IgG1 (Alpha Diagnostic International). Diarrhea assessments (profuse liquid stool) and hyperthermia measurements (rectal temperature drop > 2°C) are performed as previously described4 (link).
Lee J.B., Chen C.Y., Liu B., Mugge L., Angkasekwinai P., Facchinetti V., Dong C., Liu Y.J., Rothenberg M.E., Hogan S.P., Finkelman F.D, & Wang Y.H. (2015). IL-25 and CD4+TH2 cells enhance ILC2-derived IL-13 production that promotes IgE-mediated experimental food allergy. The Journal of allergy and clinical immunology, 137(4), 1216-1225.e5.
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6

Quantification of Mast Cell Markers

Cited 1 time
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Levels of Mcpt1 (eBioscience, San Diego, CA), Mcpt4 (Aviva Systems Biology, San Diego, CA), and IgE (eBioscience) were determined in plasma samples using commercial ELISAs and a microplate reader (BMG LABTECH, Cary, NC). IL-33 was measured in ileum tissue (prepared as in Ref. 17 (link)) using a Bio-Plex Pro Mouse Cytokine IL-33 Set (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions.
Donnelly E.L., Céspedes N., Hansten G., Wagers D., Briggs A.M., Lowder C., Schauer J., Haapanen L., Van de Water J, & Luckhart S. (2022). The Basophil IL-18 Receptor Precisely Regulates the Host Immune Response and Malaria-Induced Intestinal Permeability and Alters Parasite Transmission to Mosquitoes without Effect on Gametocytemia. ImmunoHorizons, 6(8), 630-641.
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7

Quantification of Mast Cell Markers

Cited 1 time
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Levels of Mcpt1 (eBioscience, San Diego, CA), Mcpt4 (Aviva Systems Biology, San Diego, CA), and IgE (eBioscience) were determined in plasma samples using commercial ELISAs and a microplate reader (BMG LABTECH, Cary, NC). IL-33 was measured in ileum tissue (prepared as in Ref. 17 (link)) using a Bio-Plex Pro Mouse Cytokine IL-33 Set (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions.
Donnelly E.L., Céspedes N., Hansten G., Wagers D., Briggs A.M., Lowder C., Schauer J., Haapanen L., Van de Water J, & Luckhart S. (2022). The Basophil IL-18 Receptor Precisely Regulates the Host Immune Response and Malaria-Induced Intestinal Permeability and Alters Parasite Transmission to Mosquitoes without Effect on Gametocytemia. ImmunoHorizons, 6(8), 630-641.
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8

Quantifying Plasma Immune Markers

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Levels of plasma IgE (eBioscience, San Diego, CA; Thermo Fisher Scientific, Waltham, MA), Mcpt4 (Aviva Systems Biology, San Diego, CA), Mcpt1 (eBioscience), and histamine (Enzo Life Sciences, Farmingdale, NY) were determined in plasma samples using commercial ELISAs according to the manufacturers’ instructions and a microplate reader (BMG Labtech, Cary, NC).
Donnelly E.L., Céspedes N., Hansten G., Wagers D., Briggs A.M., Lowder C., Schauer J., Garrison S.M., Haapanen L., Van de Water J, & Luckhart S. (2022). Basophil Depletion Alters Host Immunity, Intestinal Permeability, and Mammalian Host-to-Mosquito Transmission in Malaria. ImmunoHorizons, 6(8), 581-599.
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9

Murine Food Allergy Evaluation

Cited 1 time
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Serum samples were analyzed using ELISA kits for OVA-specific IgE (MD Bioproducts), MCPt-1 (eBioscience), and OVA-specific IgG1 (alpha diagnostic international). For intestinal histological analyses, duodenal tissue was fixed in 10% formalin and processed by standard histological techniques. 5-μm tissue sections were stained with Leder stain for chloroacetate esterase (CAE) activity in intestinal mast cells or periodic acid-Schiff (PAS) for mucins in goblet cells. Stained cells were quantified as previously described (Brandt et al., 2003 (link)). Assessment of diarrhea and measurement of hyperthermia (rectal temperature drop > 2°C) were performed as previously described (Osterfeld et al., 2010 (link)).
Chen C.Y., Lee J.B., Liu B., Ohta S., Wang P.Y., Kartashov A., Mugge L., Abonia J.P., Barski A., Izuhara K., Rothenberg M.E., Finkelman F.D., Hogan S.P, & Wang Y.H. (2015). Induction of Interleukin-9-producing Mucosal Mast cells Promotes Susceptibility to IgE-mediated Experimental Food Allergy. Immunity, 43(4), 788-802.
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10

Quantifying Plasma Immune Markers

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Levels of plasma IgE (eBioscience; Thermo Fisher Scientific, Inc.), Mcpt1 (eBioscience) neutrophil elastase (NE) (Abcam) and myeloperoxidase (MPO) (eBioscience; Thermo Fisher Scientific, Inc.) were determined for individual mice in the three replicates (n=109) using commercial ELISAs according to manufacturer’s instructions and a microplate reader (Molecular Devices, LLC).
Céspedes N., Donnelly E.L., Lowder C., Hansten G., Wagers D., Briggs A.M., Schauer J., Haapanen L., Åbrink M., Van de Water J, & Luckhart S. (2022). Mast Cell Chymase/Mcpt4 Suppresses the Host Immune Response to Plasmodium yoelii, Limits Malaria-Associated Disruption of Intestinal Barrier Integrity and Reduces Parasite Transmission to Anopheles stephensi. Frontiers in Immunology, 13, 801120.
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