human osteopontin spp1 elisa kit  (Boster Bio)

Journal: Bioengineering

Article Title: In Vivo Assessment of Laboratory-Grown Kidney Tissue Grafts

doi: 10.3390/bioengineering10111261

Figure Lengend Snippet: Identification of CD59 as a human-specific biomarker for the proximal nephron. ( A ). Schematic of the screening approach: normal human urinary proteome, in vitro cultured kidney organoid transcriptome, and transcriptome of engrafted kidney organoids were cross-referenced for common proteins/transcripts. ( B ). Amino acid identities of human versus mouse paralogs and results of measurement of human versus mouse using commercial ELISA kits. ( C ). Expression of SPP1 in adult human kidney. ( D ). Expression of CD59 in adult human kidney. ( E ). Results of ELISA measurement of 3 male and 3 female urine samples from humans and mice. #NUM! indicates a measurement below the threshold of detection, 3.21 × 10 4 is the Glomax plate reader convention for 3.21 × 10 4 . ( F ). Schematic showing the origin of CD59 secretion (green) and uromodulin (THP secretion (purple).

Article Snippet: Assays for uromodulin (Human Uromodulin DuoSet ELISA DY5144-05, R&D Systems, Minneapolis, MN, USA), CD59 (Human CD59 ELISA ab263893, Abcam, Waltham, MA, USA), PSAP (Human PSAP/Prosaposin (Sandwich ELISA) ELISA LS-F35235, Lifespan Biosciences, Lynnwood, MA, USA), cubilin (Human CUBN/Cubilin (Sandwich ELISA) ELISA LS-F38077, Lifespan Biosciences, Lynnwood, MA, USA), SPP1 (Human Osteopontin (OPN) Quantikine ELISA DOST00, R&D Systems, Minneapolis, MN, USA), and HSPG2 (HSPG2 elisa kit: Human Basement membrane-specific heparan sulfate proteoglycan core protein ELISA MBS765938, MyBioSource, San Diego, CA, USA) were used according to the manufacturer’s instructions and assays were read on a Glomax Explorer (Promega, Madison, WI, USA) plate reader.

Techniques: Biomarker Discovery, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Molecular Oncology

Article Title: Hypoxia‐driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial–mesenchymal transition and cancer stem cell‐like properties by modulating forkhead box protein M1

doi: 10.1002/1878-0261.12399

Figure Lengend Snippet: Hypoxia‐driven ROS ‐induced PSC activation increases the expression and secretion of OPN . (A) Subconfluent PSC s were cultured in normoxia or hypoxia with or without NAC , and then, the mRNA expression of OPN was analyzed by quantitative real‐time RT ‐ PCR . (B and C) PSC s were treated as in (A), and the protein expression of OPN was determined by western blot analysis. β‐Actin was used as an internal control. (D) PSC s were treated as in (A), and the OPN secretion was evaluated by ELISA . All data are shown as the mean ± SD of at least three independent experiments. One‐way ANOVA and Tukey's multiple comparisons test were performed to determine significance. * P < 0.05, ** P < 0.01.

Article Snippet: The production of OPN in the supernatants was quantified using a human OPN ELISA kit (Boster, Wuhan, China) according to the manufacturer's recommendations.

Techniques: Activation Assay, Expressing, Cell Culture, Quantitative RT-PCR, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Molecular Oncology

Article Title: Hypoxia‐driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial–mesenchymal transition and cancer stem cell‐like properties by modulating forkhead box protein M1

doi: 10.1002/1878-0261.12399

Figure Lengend Snippet: Hypoxia‐driven paracrine OPN signaling promotes EMT and CSC ‐like properties in PCC s. (A) The mRNA expression levels of OPN , the αv subunit of integrin, and the β3 subunit of integrin in 4 pancreatic cancer cell lines and PSC s were analyzed by quantitative real‐time RT ‐ PCR . (B and C) The protein expression levels of OPN , the αv subunit of integrin, and the β3 subunit of integrin in 4 pancreatic cancer cell lines and PSC s were determined by western blot analysis. β‐Actin was used as an internal control. (D) The cell supernatants of 4 pancreatic cancer cell lines and PSC s were collected, and the OPN concentration was measured by ELISA . (E) Representative images from the wound healing assay after rh OPN , PSC ‐ CM , or PSC ‐ CM + OPN Ab treatments in Bx PC ‐3 and CFPAC ‐1 cells. Images were obtained at 0 h and 24 h. The scale bar represents 100 μm. (F and G) Representative images of the Matrigel invasion assay after rh OPN , PSC ‐ CM , or PSC ‐ CM + OPN Ab treatments in Bx PC ‐3 and CFPAC ‐1 cells. The invasive cells were quantified by counting the number of cells in 10 random fields. The scale bar represents 50 μm. (H, I and J) Representative images of the tumorsphere formation assay after rh OPN , PSC ‐ CM , or PSC ‐ CM + OPN Ab treatments in Bx PC ‐3 and CFPAC ‐1 cells. The number of tumorspheres was counted and plotted, and the percentage of tumorspheres with diameters of 50–100 μm, 100–150 μm, or >150 μm was calculated and plotted. The scale bar represents 50 μm. (K) The protein expression levels of EMT and CSC markers after rh OPN , PSC ‐ CM , or PSC ‐ CM + OPN Ab treatments were determined by western blot analysis in Bx PC ‐3 and CFPAC ‐1 cells. β‐Actin was used as an internal control. Q‐ PSC : Quiescent pancreatic stellate cell. A‐ PSC : Activated pancreatic stellate cell. All data are shown as the mean ± SD of at least three independent experiments. One‐way ANOVA and Tukey's multiple comparisons test were performed to determine significance. * P < 0.05, ** P < 0.01, ns: no significance.

Article Snippet: The production of OPN in the supernatants was quantified using a human OPN ELISA kit (Boster, Wuhan, China) according to the manufacturer's recommendations.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Invasion Assay, Tube Formation Assay