Serpine1

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

SERPINE1 is a protein-coding gene that produces the plasminogen activator inhibitor-1 (PAI-1) protein. PAI-1 is a serine protease inhibitor that regulates the fibrinolytic system by inhibiting tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA).

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Lab products found in correlation

Power sybr green, by Thermo Fisher Scientific (2 mentions) Abi 7900, by Thermo Fisher Scientific (2 mentions) Sybr green, by Thermo Fisher Scientific (2 mentions) Superscript first strand kit, by Thermo Fisher Scientific (2 mentions) Prism software, by GraphPad (1 mentions) 7300 real time pcr system, by Thermo Fisher Scientific (1 mentions) Eomes, by Thermo Fisher Scientific (1 mentions) Advantage rt for pcr kit, by Takara Bio (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions) Rneasy extraction kit, by Qiagen (1 mentions) Human tnf, by Merck Group (1 mentions)

5 protocols using serpine1

Quantitative Real-Time PCR for Gene Expression

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All RNA samples were treated with DNase to remove any contaminant genomic DNA. RNA was then reverse transcribed using Superscript first strand kit (Life technologies) and the cDNA was diluted and used as template for SYBR Green or TaqMan qPCR on the ABI 7900 (Life technologies). RNA from human brain, heart, kidney, liver, lung and muscle was purchased from life technologies. Human neurons and astrocytes RNA have been purchased from ScienceCell research laboratories, cat# 1525 and cat# 1585 respectively. SERPINE1, TAC1, PGK1 and b-ACTIN TaqMan assays were purchased from Life Technologies. Primers used for SYBR Green qRT-PCR to validate and measure the expression of novel ncRNAs are listed in Additional File 7. Non-coding RNAs expression was measured using Power SYBR Green (Life Technologies). For all qRT-PCR reactions, we included three technical replicates. To compare the expression of genes across different cellular compartment, GraphPad prism software was used to perform ANOVA followed by Tukey post-hoc test. A p value of below 0.05 was considered as statistically significant. The Student’s t-test was used to compare the expression between the normal brain and LOAD.

Magistri M., Velmeshev D., Makhmutova M, & Faghihi M.A. (2015). Transcriptomics Profiling of Alzheimer’s Disease Reveal Neurovascular Defects, Altered Amyloid-β Homeostasis, and Deregulated Expression of Long Noncoding RNAs. Journal of Alzheimer's Disease, 48(3), 647-665.

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Quantification of ncRNA Expression in Human Tissues

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All RNA samples were treated with DNase to remove any contaminant genomic DNA. RNA was then reverse transcribed using Superscript first strand kit (Life Technologies) and the cDNA was diluted and used as template for SYBR Green or TaqMan qPCR on the ABI 7900 (Life Technologies). RNA from human brain, heart, kidney, liver, lung, and muscle was purchased from Life Technologies. Human neurons and astrocytes RNA have been purchased from ScienceCell Research Laboratories, cat# 1525 and cat# 1585 respectively. SERPINE1, TAC1, PGK1 and β-ACTIN TaqMan assays were purchased from Life Technologies. Primers used for SYBR Green qRT-PCR to validate and measure the expression of novel ncRNAs are listed in Supplementary File 7. Non-coding RNAs expression was measured using Power SYBR Green (Life Technologies). For all qRT-PCR reactions, we included three technical replicates. To compare the expression of genes across different cellular compartment, GraphPad prism software was used to perform ANOVA followed by Tukey post-hoc test. A p value of below 0.05 was considered as statistically significant. The Student’s t-test was used to compare the expression between the normal brain and LOAD.

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Quantitative RT-PCR Analysis of NK Cells

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Total RNA was isolated from the NK cells using the RNeasy Extraction Kit (Qiagen, Valencia, CA). RNA was reverse transcribed into cDNA using the Advantage RT-for-PCR Kit (Clontech, Mountain View, CA). Resultant cDNA (100 ng) was quantitated using TaqMan primers and probes as follows: CD226 (Hs00170832_m1), KLRK1 (Hs00183683_m1), NCR3 (Hs01553309_g1), GZMB (Hs00188051_m1), PRF1 (Hs00169473_m1), EOMES (Hs00172872_m1), JUN (Hs01103582_s1), SERPINE1 (Hs00167155_m1), SMAD7 (Hs00998193_m1), and GAPDH (Hs02786624_g1) (Life Technologies, Grand Island, NY). Each mRNA expression level was calculated as expression relative to GAPDH. Real-time PCR was performed on the 7300 Real-Time PCR System (Applied Biosystems, Carlsbad, CA).

Fujii R., Jochems C., Tritsch S.R., Wong H.C., Schlom J, & Hodge J.W. (2018). An IL-15 superagonist/IL-15Rα fusion complex protects and rescues NK cell cytotoxic function from TGF-β1-mediated immunosuppression. Cancer immunology, immunotherapy : CII, 67(4), 675-689.

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Tube Formation Assay on Matrigel

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Matrigel was melted at 4℃ overnight and diluted with serum-free medium (1:4). After Matrigel polymerization, human umbilical vascular endothelial cells (HUVECs, 5 × 10⁴) were suspended in a medium and seeded on the Matrigel in the 24-well plate or the lower chamber of Transwell. Human recombinant Serpin E1 (PeproTech, USA) was added to the 24-well plate at a concentration of 2 ng/ml, or 1 × 10⁵ cells, including AGS, MNK45, and primary gastric cancer cells overexpressing Serpin E1 and control vector, were added to the upper chamber of Transwell. After culturing for 24 h, tube formation of HUVECs was observed and photographed.

Chen X., Chen W., Zhao Y., Wang Q., Wang W., Xiang Y., Yuan H., Xie Y, & Zhou J. (2022). Interplay of Helicobacter pylori, fibroblasts, and cancer cells induces fibroblast activation and serpin E1 expression by cancer cells to promote gastric tumorigenesis. Journal of Translational Medicine, 20, 322.

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Cytokine-Mediated MMP-9 and IL-8 Expression

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Following a starvation phase overnight, 2 × 10⁶ THP-1 cells/well were stimulated with 5 ng/mL MIF, 5 ng/mL IL-13, 5 ng/mL IL-16, 5 ng/mL sICAM-1, 25 ng/mL Serpin E1, or 5 ng/mL IL-8 (Peprotech, Hamburg, Germany), either individually or in combination, for 24 h. The respective concentrations were derived from the respective product sheet by selecting a low effective concentration and then validated by dose response experiments. As a positive control, starved THP-1 cells were stimulated for 2 h with 25 ng/mL human TNF (Sigma Aldrich). Following stimulation, cells were harvested and the MMP-9 or IL-8 mRNA expression was measured.

Huber R., Attili/Abedalkhader R., Küper D., Hauke L., Lüns B., Brand K., Weissenborn K, & Lichtinghagen R. (2019). Cellular and Molecular Effects of High-Molecular-Weight Heparin on Matrix Metalloproteinase 9 Expression. International Journal of Molecular Sciences, 20(7), 1595.

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