mouse mmp13 elisa kit  (Bioss)

Journal: Journal of Pharmaceutical Analysis

Article Title: A novel exploration of COL11A1 's role in regulating myeloid-derived suppressor cell activation within the colon cancer microenvironment

doi: 10.1016/j.jpha.2024.101181

Figure Lengend Snippet: Effects of collagen type XI alpha 1 ( COL11A1 ) on growth, proliferation, and migration of cancer-associated fibroblasts (CAFs). (A) COL11A1 , matrix metalloproteinase ( MMP )3, and MMP13 messenger RNA (mRNA) expression in normal fibroblasts (NFs) and CAFs. (B) Protein expression of COL11A1, MMP3, and MMP13 in NFs and CAFs. (C) Enzyme-linked immunosorbent assay (ELISA) detection of MMP3 and MMP13 content in supernatant of bone marrow cells (BMCs) co-cultured with NFs and CAFs. (D) mRNA expression levels of COL11A1 after knockout. (E) Protein expression levels of COL11A1 after knockout. (F) Cell counting kit (CCK)-8 assay to evaluate the growth activity of CAFs after COL11A1 knockout ( COL11A1 KO ). (G) 5-Ethynyl-2′-deoxyuridine (EdU) labeling to measure the proliferation rate of CAFs after COL11A1 KO . (H) Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining to assess the apoptosis rate of CAFs after COL11A1 KO . (I) Wound healing experiment to determine the migration ability of CAFs after COL11A1 KO . (J) Expression of relevant proteins in CAFs after COL11A1 KO . ∗∗ P < 0.01; all cell experiments were repeated three times. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; OD: optical density; DAPI: 4′,6-diamidino-2-phenylindole; p-PI3K: phosphorylated-phosphoinositide 3-kinase (PI3K); p-AKT: phosphorylated-protein kinase B (AKT); p-mTOR: phosphorylated mechanistic target of rapamycin (mTOR).

Article Snippet: The levels of matrix metalloproteinase (MMP)3 and MMP13 in the cell supernatant were detected using the mouse MMP3 ELISA kit (D721078, Bioss, Beijing, China) and mouse MMP13 ELISA kit (D721035, Bioss, Beijing, China).

Techniques: Migration, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Knock-Out, Cell Counting, CCK-8 Assay, Activity Assay, Labeling, TUNEL Assay, Staining

Journal: Journal of Pharmaceutical Analysis

Article Title: A novel exploration of COL11A1 's role in regulating myeloid-derived suppressor cell activation within the colon cancer microenvironment

doi: 10.1016/j.jpha.2024.101181

Figure Lengend Snippet: Effects of collagen type XI alpha 1 ( COL11A1 ) in cancer-associated fibroblasts (CAFs) on myeloid-derived suppressor cells (MDSCs) differentiation and activation. (A) Enzyme-linked immunosorbent assay (ELISA) detection of matrix metalloproteinase (MMP)3 and MMP13 content in supernatant of bone marrow cells (BMCs) co-cultured with COL11A1 knockout ( COL11A1 KO ) CAFs. (B) Flow cytometry analysis of arginase 2 ( ARG2 ) and CD11b expression frequency in BMCs co-cultured with COL11A1 KO CAFs. (C) ARG2 and CD11b messenger RNA (mRNA) expression levels in BMCs co-cultured with COL11A1 KO CAFs. (D) ARG2 and CD11b protein expression levels in BMCs co-cultured with COL11A1 KO CAFs. (E) Flow cytometry analysis of interferon gamma (IFN-γ) secretion in CD8 + T cells. (F) Flow cytometry analysis of granzyme B (GZMB) secretion in CD8 + T cells. (G) Flow cytometry analysis of tumor necrosis factor alpha (TNF-α) secretion in CD8 + T cells. (H) Flow cytometry analysis of apoptosis in CD8 + T cells. (I) Flow cytometry analysis of proliferation in CD8 + T cells. ∗∗ P < 0.01; all cell experiments were repeated three times. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; FITC: fluorescein isothiocyanate; CFSE: carboxyfluorescein succinimidyl ester.

Article Snippet: The levels of matrix metalloproteinase (MMP)3 and MMP13 in the cell supernatant were detected using the mouse MMP3 ELISA kit (D721078, Bioss, Beijing, China) and mouse MMP13 ELISA kit (D721035, Bioss, Beijing, China).

Techniques: Derivative Assay, Activation Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Knock-Out, Flow Cytometry, Expressing

Journal: Journal of Pharmaceutical Analysis

Article Title: A novel exploration of COL11A1 's role in regulating myeloid-derived suppressor cell activation within the colon cancer microenvironment

doi: 10.1016/j.jpha.2024.101181

Figure Lengend Snippet: Impact of collagen type XI alpha 1 ( COL11A1 ) activation through myeloid-derived suppressor cells (MDSCs) on colon cancer (CC) tumor growth and proliferation. (A) Growth of CC orthotopic xenografts in mice after COL11A1 knockout. (B) Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining to evaluate cell apoptosis in CC orthotopic xenografts in mice after COL11A1 knockout. (C) Ki67 staining to assess cell proliferation in CC orthotopic xenografts in mice after COL11A1 knockout. (D) Immunohistochemical detection of COL11A1 and arginase 2 ( ARG2 ) expression in CC orthotopic xenografts in mice after COL11A1 knockout. (E) Immunohistochemical detection of matrix metalloproteinase ( MMP )3 and MMP13 expression in CC orthotopic xenografts in mice after COL11A1 knockout. (F) Flow cytometry analysis of MDSC differentiation and activation in CC orthotopic xenografts in mice after COL11A1 knockout ( n = 6 per group). ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: The levels of matrix metalloproteinase (MMP)3 and MMP13 in the cell supernatant were detected using the mouse MMP3 ELISA kit (D721078, Bioss, Beijing, China) and mouse MMP13 ELISA kit (D721035, Bioss, Beijing, China).

Techniques: Activation Assay, Derivative Assay, Knock-Out, TUNEL Assay, Staining, Immunohistochemical staining, Expressing, Flow Cytometry

Journal: Journal of Pharmaceutical Analysis

Article Title: A novel exploration of COL11A1 's role in regulating myeloid-derived suppressor cell activation within the colon cancer microenvironment

doi: 10.1016/j.jpha.2024.101181

Figure Lengend Snippet: Collagen type XI alpha 1 ( COL11A1 ) induces the paracrine secretion of matrix metalloproteinase (MMP)3 and MMP13 in cancer-associated fibroblasts (CAFs), promoting colon cancer (CC) development and immune escape (Created by using www.biorender.com ).

Article Snippet: The levels of matrix metalloproteinase (MMP)3 and MMP13 in the cell supernatant were detected using the mouse MMP3 ELISA kit (D721078, Bioss, Beijing, China) and mouse MMP13 ELISA kit (D721035, Bioss, Beijing, China).

Techniques:

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Anti-inflammatory role of Cuprorivaite (CaCuSi 4 O 10 ) microspheres in IL-1β-stimulated chondrocytes. (A) Cell counting kit-8 test for cell proliferation. (B) Fluorescein diacetate for cell viability detection. (C) Cell apoptosis by flow cytometry. (D) ELISA detection for TNF-α, IL-6 and MMP13 in cell supernatant. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cell lysates. (F) RT-qPCR quantification for in TNF-α, IL-6 and MMP13. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Cell Counting, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Cuprorivaite (CaCuSi 4 O 10 ) microspheres improved IL-1β-induced injury in chondrocytes via inhibiting Wnt/β-catenin pathway. (A) Cell counting kit-8 test for cell viability. (B) Fluorescein diacetate for cell viability detection. (C) Cell apoptosis by flow cytometry. (D) ELISA detection for TNF-α, IL-6 and MMP13 in cell supernatant. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cell lysates. (F) RT-qPCR quantification for extracellular matrix components including collagen II and SOX9. (G) Intracellular copper content. (H) RT-qPCR quantification for cuproptosis biomarkers including ATP7B and FDX1. (I) Western blot detection for ATP7B and FDX1. (J) Oxidative stress detection including MDA, SOD and GSH. (K) RT-qPCR quantification for Wnt1, GSK3β and β-catenin. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Cell Counting, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Validation of mechanism of Cuprorivaite (CaCuSi 4 O 10 ) microspheres in OA. (A) Hematoxylin and eosin staining. (B) Safranin-O staining/fast green staining. (C) OARSI score. (D) ELISA detection for TNF-α, IL-6 and MMP13 in the serum. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cartilage tissue. (F) RT-qPCR quantification for extracellular matrix components including collagen II and SOX9 in cartilage tissue. (G) Copper content in cartilage tissue. (H) RT-qPCR quantification for cuproptosis biomarkers including ATP7B and FDX1 in cartilage tissue. (I) Western blot detection for ATP7B and FDX1 in cartilage tissue. (J) Representative images of FDX1 expression in cartilage tissue detected by immunohistochemistry. (K) Oxidative stress detection including MDA, SOD and GSH in cartilage tissue. (L) RT-qPCR quantification for Wnt1, GSK3β and β-catenin in cartilage tissue. ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Biomarker Discovery, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR, Expressing, Immunohistochemistry