Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: Label-free mass spectrometry data. List of statistically significant discovered proteins using LC-MS analysis. Proteins (in bold) were selected for further validation using ELISAs.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques: Mass Spectrometry, Biomarker Discovery, Variant Assay

Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: ELISA Data. Mean, SD, Area under the curve (AUC) and p -value for each of the new and standard proteins found in the two groups of patients compared.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: BBA Clinical

Article Title: Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients

doi: 10.1016/j.bbacli.2017.05.003

Figure Lengend Snippet: Logistic regression analysis data. List of different protein combinations used to establish the best model that can be used as a predictive panel for response to induction therapy containing bortezomib regime.

Article Snippet: Four commercially available kits for these four proteins; angiogenin (ANG) [Abcam, UK - ab99970], clusterin (CLU) [R&D system, UK - DCLU00], C-C Motif Chemokine 18 (CCL18) [Abcam, UK - ab100620], and Complement C1q [Abcam, UK - ab170246] were used.

Techniques:

Journal: Orphanet Journal of Rare Diseases

Article Title: Specific populations of urinary extracellular vesicles and proteins differentiate type 1 primary hyperoxaluria patients without and with nephrocalcinosis or kidney stones

doi: 10.1186/s13023-020-01607-1

Figure Lengend Snippet: Number of urinary extracellular vesicles (EVs) carrying cellular adhesion/inflammatory and renal injury molecules from primary hyperoxaluria type 1 patients without and with nephrocalcinosis (NC) or kidney stones (KS)

Article Snippet: PE conjugated mouse anti-human clusterin antibody purchased from Novus Biologicals, LLC, Centennial, CO, USA.

Techniques:

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 1. Clusterin expression in luminal breast cancer samples. A and B. Clusterin expression was analyzed by immunohistochemistry in tumoral (a) and juxtatumoral (b) tissues. Representative pictures are shown (n = 3, bars: 50 µm). C. Clusterin expression was quantified in tumor samples and their juxtatumoral counterparts by ELISA. Results are expressed as the amount of clusterin/total protein (µg of clusterin/mg of total protein, n = 21, ns = no statistically significant differences, p = .31).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 2. Fucosylated clusterin (fCLU) is expressed in luminal breast cancer and interacts with DC-SIGN. a. Scheme of the ELISA assay used to detect fucosylated clusterin. Clusterin is captured by a mAb directed to clusterin and the presence of fucosylated clusterin (fCLU) is revealed by using either biotinilated Ulex europaeus agglutinin-I lectin (UEA-I) or Lotus tetragonolobus lectin (LT). b. Fucosylated semen clusterin, but not serum or recombinant clusterin (blue bars), was detected by the assay described in (a). The addition of α-L-fucose prevents the recognition of fucosylated semen clusterin (red bars). Results are expressed as the mean ± SD of 4 independent experiments. c and d. Fucosylated clusterin expression in breast tumor and juxtatumoral samples (n = 14–21) was analyzed using Lotus tetragonolobus lectin (c) or Ulex europaeus agglutinin-I (d). Results are expressed as the amount of fucosylated clusterin relative to the amount of total clusterin (left panels) or total protein (right panels) (***p < .001, **p < .01, *p < .05). E. The ability of clusterin from tumor samples and juxtatumoral samples to bind to DC-SIGN was evaluated. Upper panel: the presence of clusterin was evaluated using a mAb directed to clusterin. Lower panel: the membrane was stripped and revealed using DC-SIGN-huFc and HRP conjugated anti-huIgG. Of note, total clusterin but not total protein was used to normalize the amount of sample loaded into the gel for each tumor and juxtatumor sample pair. Accordingly, no loading control was analyzed. Panel F shows the ratio between DC-SIGN-huFC binding and total CLU. The fold change between tumor and juxtatumor CLU ratios are shown.

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Expressing, Membrane, Control, Binding Assay

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 6. Fucosylated clusterin promotes the differentiation of macrophages into a proangiogenic profile. a-c. Macrophages were differentiated from monocytes cultured with M-CSF for 5 days, in the absence or presence of semen fucosylated clusterin. Then, cells were stimulated, or not, with LPS (10 ng/ml), and the pattern of cell clustering (a), the expression of HLA-DR, CD86, PDL-1, CD40 and CD80 (b), and the production of VEGF, IL-8, TNF-α, IL-10, and IL-6 (c) were evaluated by optical microscopy, flow cytometry or ELISA. Representative pictures (bars = 100µm) and histograms (n = 3–9) are shown in a and b. On b, the control condition mean fluorescent intensities (MFI) of each independent experiment were normalized as 1. The mean of 5–9 independent experiments is shown. The mean of 5–7 independent experiments is shown in c (**p < .01, *p < .05).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Cell Culture, Expressing, Microscopy, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Control

Journal: OncoImmunology

Article Title: Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN)

doi: 10.1080/2162402x.2019.1629257

Figure Lengend Snippet: Figure 5. Fucosylated clusterin interacts with macrophages. a. Expression of DC-SIGN by monocyte-derived macrophages. A representative experiment (n = 7) is shown. b. Macrophages were incubated for 30 min at 4°C with different concentrations of fucosylated clusterin isolated from semen. Cells were then washed, lysed and the binding of clusterin was revealed by ELISA. Results are expressed as the mean ± SD of 4 independent experiments. c. Macrophages were incubated for 30 min at 4°C with fucosylated clusterin (10 µg/ml), in the absence or presence of a neutralizing mAb directed to DC- SIGN (5 µg/ml), lactose (100 µg/ml), sucrose (100 µg/ml), mannan (1–100 µg/ml), or α-L-fucose (1–100 µg/ml). Cells were then washed, lysed and the binding of clusterin was revealed by ELISA. Results are expressed as the mean ± SD of 3 experiments (**p < .01, *p < .05). d. Macrophages were incubated for 15 min at 37°C with fucosylated clusterin (10 µg/ml) and clusterin endocytosis (green) was analyzed by confocal microscopy (n = 5, bars:10µm).

Article Snippet: Total clusterin was detected by ELISA using a conventional ELISA kit (Human Clusterin DuoSet ELISA, R&D Systems).

Techniques: Expressing, Derivative Assay, Incubation, Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay, Confocal Microscopy

Journal: Proteomics

Article Title: Identification, prioritization and evaluation of glycoproteins for aggressive prostate cancer using quantitative glycoproteomics and antibody-based assays on tissue specimens

doi: 10.1002/pmic.201200541

Figure Lengend Snippet: Standard Curve of ELISA assays for protein quantification of cartilage oligomeric matrix protein (COMP), periostin, membrane primary amine oxidase (VAP-1) and cathepsin L. A), dose-response curves for COMP. B), dose-response curve for periostin. C), dose-response curve for VAP-1. D), dose-response curve for cathepsin L.

Article Snippet: Materials Hydrazide resin and sodium periodate were from Bio-Rad (Hercules, CA); sequencing-grade trypsin and TMB reagent were from Promega (Madison, WI); PNGase F was from New England Biolabs (Ipswich, MA); C18 columns were from Waters (Milford, MA); Recombinant protein, capture and detection antibody of human cartilage oligomeric matrix protein (COMP), periostin, cathepsin L, clusterin and galectin-3 binding protein, streptavidin-HRP conjugates, ELISA plates and human VAP-1 Quantikine ELISA Kit were from R&D systems (Minneapolis, MN); All other chemicals were from Sigma-Aldrich (St. Louis, MO).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Proteome Science

Article Title: Development of reverse phase protein microarrays for the validation of clusterin, a mid-abundant blood biomarker

doi: 10.1186/1477-5956-7-15

Figure Lengend Snippet: Scanned image of fragments of RPPMs showing spotted triplicates of plasma and mouse IgGs probed with (+) or without (-) primary anti-clusterin antibody, and with secondary fluorescently labeled anti-IgG antibody . Pseudo-color scale, dark blue to white corresponds to increasing fluorescence.

Article Snippet: For the measurement of clusterin we used a Human Clusterin ELISA kit (BioVendor, Czech Republic).

Techniques: Labeling, Fluorescence

Journal: Proteome Science

Article Title: Development of reverse phase protein microarrays for the validation of clusterin, a mid-abundant blood biomarker

doi: 10.1186/1477-5956-7-15

Figure Lengend Snippet: Evaluation parameters of RPPMs . (a) Detection of clusterin in plasma samples using monoclonal and (b) polyclonal primary anti-clusterin antibodies. The log 10 average signal intensity for clusterin was plotted against plasma dilution 1/4 to 1/512. (c) Minimum difference in clusterin concentration detected on RPPMs. Twenty different concentrations of recombinant clusterin were spiked in plasma and the average fluorescence intensity of three slides median normalized was plotted against the ten highest clusterin concentrations. The average fluorescence level (▲) with the value of 2 standard deviations (---) for the ten lowest endogenous clusterin concentrations are used as background clusterin level in this experiment. Arrow points to the concentration of spiked clusterin that yielded a signal at least 2SD above background plasma clusterin. (d) Correlation of clusterin levels measured with ELISA and RPPM (r = 0.989). Clusterin was measured in plasma samples spiked with increasing concentrations (0.9–500 μg/ml) of recombinant clusterin.

Article Snippet: For the measurement of clusterin we used a Human Clusterin ELISA kit (BioVendor, Czech Republic).

Techniques: Concentration Assay, Recombinant, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Proteome Science

Article Title: Development of reverse phase protein microarrays for the validation of clusterin, a mid-abundant blood biomarker

doi: 10.1186/1477-5956-7-15

Figure Lengend Snippet: Scanned image of slide containing 149 clinical samples spotted in quadruplicate and probed for clusterin .

Article Snippet: For the measurement of clusterin we used a Human Clusterin ELISA kit (BioVendor, Czech Republic).

Techniques:

Journal: Proteome Science

Article Title: Development of reverse phase protein microarrays for the validation of clusterin, a mid-abundant blood biomarker

doi: 10.1186/1477-5956-7-15

Figure Lengend Snippet: Measurement of clinical samples using RPPMs andELISA . (a) The log 10 median clusterin intensity of all clinical samples analysed was plotted. (b) Clusterin levels in serum and CTAD plasma samples taken from the same individual and processed at different time points (0 min to 24 hrs) were measured with ELISA and (c) RPPM. (d) Correlation of RPPM data and ELISA values obtained for all clinical samples. The scaled data of all 16 data points per sample was averaged and then divided by its respective clusterin concentration measured by ELISA to obtain a ratio. For each sample, the log 2 ratio was plotted against its respective clusterin concentration measured by ELISA.

Article Snippet: For the measurement of clusterin we used a Human Clusterin ELISA kit (BioVendor, Czech Republic).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay