human dkk 1 duoset elisa kit  (R&D Systems)

Journal: Cancer Medicine

Article Title: Dickkopf 1 is expressed in normal fibroblasts during early stages of colorectal tumorigenesis

doi: 10.1002/cam4.6992

Figure Lengend Snippet: Identified 18 overexpressed genes.

Article Snippet: DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems) and Human DKK‐1 DuoSet ELISA Kit (R&D Systems) were used for the experiments, and absorbance was measured using SpectraMax M2e (Molecular Devices).

Techniques:

Journal: Cancer Medicine

Article Title: Dickkopf 1 is expressed in normal fibroblasts during early stages of colorectal tumorigenesis

doi: 10.1002/cam4.6992

Figure Lengend Snippet: Immunohistochemical staining of DKK1 in clinical specimens. (A) Representative images of DKK1 in human liver tissue, DKK2 in human hepatocarcinoma tissue, SFRP1 in human heart muscle tissue in positive control (scale bar, 1000 μm), normal epithelium (scale bar, 50 μm), nonadvanced adenoma (scale bar, 200 μm), advanced adenoma (scale bar, 500 μm), and advanced cancer (scale bar, 500 μm). (B) Comparison of specimens with immunohistochemical staining for αSMA and DKK1 in (a) nonadvanced adenoma and (b) advanced cancer. Scale bars: 100 μm. DKK1 , Dickkopf‐1; αSMA , α‐smooth muscle actin.

Article Snippet: DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems) and Human DKK‐1 DuoSet ELISA Kit (R&D Systems) were used for the experiments, and absorbance was measured using SpectraMax M2e (Molecular Devices).

Techniques: Immunohistochemical staining, Staining, Positive Control, Comparison

Journal: Cancer Medicine

Article Title: Dickkopf 1 is expressed in normal fibroblasts during early stages of colorectal tumorigenesis

doi: 10.1002/cam4.6992

Figure Lengend Snippet: Histological expression of DKK1 in resection specimens.

Article Snippet: DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems) and Human DKK‐1 DuoSet ELISA Kit (R&D Systems) were used for the experiments, and absorbance was measured using SpectraMax M2e (Molecular Devices).

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Dickkopf 1 is expressed in normal fibroblasts during early stages of colorectal tumorigenesis

doi: 10.1002/cam4.6992

Figure Lengend Snippet: Functional evaluation of DKK1 using human colon cancer cell line HCT116. (A) OD values obtained using the MTS assay after 24, 48, and 72 h of incubation of HCT116 cells with recombinant human DKK1 protein. n = 6. (B) Cell invasion assay after incubation of HCT116 cells with recombinant human DKK1 protein. n = 6. (C) Relative expression of DKK1 after siRNA transfection using Lipofectamine 3000. (D) MTS assay in HCT116 cells at 24, 48, and 72 h after siRNA transfection. n = 4. * p < 0.05; ** p < 0.01; *** p < 0.001. Rec DKK1 = recombinant human DKK1 protein.

Article Snippet: DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems) and Human DKK‐1 DuoSet ELISA Kit (R&D Systems) were used for the experiments, and absorbance was measured using SpectraMax M2e (Molecular Devices).

Techniques: Functional Assay, MTS Assay, Incubation, Recombinant, Invasion Assay, Expressing, Transfection

Journal: Cancer Medicine

Article Title: Dickkopf 1 is expressed in normal fibroblasts during early stages of colorectal tumorigenesis

doi: 10.1002/cam4.6992

Figure Lengend Snippet: Serum DKK1 expression analysis. (A, B) ELISA of supernatant during co‐culture at Days 5–7; n = 2. Left: NFs monoculture, right: NFs co‐cultured with organoid (A) Using adenoma organoid. (B) Using cancer organoid. (C–F) Serum DKK1 expression in patients with colorectal tumors. (C) Comparison of serum DKK1 in samples obtained from patients with advanced cancer ( n = 32) and control ( n = 17). (D) ROC curve for (C), AUC = 0.78. (E) Comparison of serum DKK1 in paired serum samples from patients with advanced adenoma before and after treatment ( n = 17). (F) ROC curve for (E), AUC = 0.64. In (C) and (D), * p < 0.05; *** p < 0.001. AUC, area under the curve; DKK1 , Dickkopf‐1; post, post‐treatment; pre, pre‐treatment; ROC, receiver operating characteristic.

Article Snippet: DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems) and Human DKK‐1 DuoSet ELISA Kit (R&D Systems) were used for the experiments, and absorbance was measured using SpectraMax M2e (Molecular Devices).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Cell Culture, Comparison, Control

Journal: Alzheimer's & Dementia

Article Title: Amyloid β synaptotoxicity is Wnt-PCP dependent and blocked by fasudil

doi: 10.1016/j.jalz.2017.09.008

Figure Lengend Snippet: Aβ synaptotoxicity is Dkk1 dependent. (A) Rat primary cortical neuronal cultures (14 d.i.v.) were treated with 10 μM Aβ 25–35 for 2 and 3 hours, cells were harvested for RNA extraction, and media were collected for protein analysis. cDNA was generated and qRT-PCR performed to determine rat Dkk1 mRNA levels, left. Secreted Dkk1 protein levels in media were measured by ELISA, right. (B) Similar cultures were treated at 3 μM and 300 nM with AβO preparation for the times indicated and harvested, and Dkk1 mRNA levels were determined as mentioned previously. (C and D) Similar cultures were transfected with eGFP at 24 d.i.v., 48 hours later treated for 4 hours with 2 μM AβO, or for 3 hours with 400 ng/mL Dkk1, fixed, imaged by confocal microscopy, and dendritic spine density and morphology assessed. Both treatments resulted in a significant reduction in dendritic spine linear density, quantified in (D), scale bar = 5 μM. Dendritic spine linear density/10 μm: control, 5.8 ± 0.41; AβO, 3.7 ± 0.34; Dkk1, 3.5 ± 0.31; P < .001 for all treatments. (E and F) Similar cultures were treated overnight with Dkk1-siRNA duplex, or a scrambled version as control, each linked to the Pen-1 peptide. Next day, cells were treated with 2 μM AβO for 4 hours, fixed, and fluorescently labeled with phalloidin-488, imaged (E), and F-actin–labeled puncta quantified (F), scale bar = 50 μM. In all the aforementioned, significance was determined by ANOVA and Tukey's post hoc t-test. Error bars indicate standard error of the mean. Abbreviations: AβO, amyloid β (1–42) oligomers; ANOVA, analysis of variance; d.i.v., days in vitro; eGFP, enhanced green fluorescent protein; Dkk1, Dickkopf-1.

Article Snippet: Rat Dkk1 mRNA expression was performed by quantitative reverse transcription-polymerase chain reaction and protein levels determined using a DuoSet ELISA kit (DY1906; R&D Systems), both as previously described .

Techniques: RNA Extraction, Generated, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Transfection, Confocal Microscopy, Control, Labeling, In Vitro