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: Human reproduction (Oxford, England)

Article Title: CDC42 deficiency leads to endometrial stromal cell senescence in recurrent implantation failure.

doi: 10.1093/humrep/deae246

Figure Lengend Snippet: Figure 1. Cellular senescence in the stroma of recurrent implantation (RIF) failure endometrium. (A) Gene ontology (GO) biological process enrichment of the differentially expressed genes (DEGs) between fertile control (CTR) patients and patients with RIF in SRP224538. (B) FPKM level of TP53 in CTR patients and RIF patients in SRP224538. (C) FPKM level of IL6 in CTR patients and RIF patients in SRP224538. (D–F) Levels of IL6, CLU, and sST2 secretion in primary endometrial stromal cell supernatant from CTR and RIF patients before and after 72 h of 8Br-cAMPþMPA treatment. (G and H) Representative images of immunohistochemical (IHC) staining of P16 in mid-secretory endometrium of CTR patients (n ¼ 22) versus RIF patients (n ¼ 22). Black arrows indicate typical P16þ cells. Quantitative analysis by number of P16þ cells per 200× field for P16 staining. (I and J) Representative images of IHC staining of P21 in mid-secretory endometrium of CTR patients (n ¼ 22) versus RIF patients (n ¼ 22). Black arrows indicate typical P21þ

Article Snippet: Commercially available ELISA kits were utilized to measure the level of secreted protein (PRL: Elabscience, No. E-EL-H0141; IGFBP1: Elabscience, No. E-EL-H0442; IL6: Elabscience, No. E-EL-H6156; IL8: MULTI SCIENCES, No. EK10896; CLU: Elabscience, No. E-EL-H0038; sST2: Elabscience, No. E-EL-H6082) in collected supernatants according to the manufacturer’s instructions.

Techniques: Control, Immunohistochemical staining, Immunohistochemistry, Staining

Journal: Human reproduction (Oxford, England)

Article Title: CDC42 deficiency leads to endometrial stromal cell senescence in recurrent implantation failure.

doi: 10.1093/humrep/deae246

Figure Lengend Snippet: Figure 2. CDC42 knockdown accelerates senescence of differentiated endometrial stromal cells (EnSCs). (A–C) Expression of CDC42, CDKN2A, and CDKN1A mRNA levels in primary EnSCs after 72 h of adenovirus treatment. (D) Expression of CDC42, P21, P53, and p-γ-H2AX protein levels in primary EnSCs after 72 h of adenovirus treatment. (E) Expressions of collagen I, collagen III, collagen IV, and MMP2 protein levels in primary EnSC after 72 h of adenovirus treatment. (F–H) Expressions of CDC42, CDKN2A, and CDKN1A mRNA levels in primary EnSC transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (I–K) Level of IL6, CLU, and sST2 secretion in primary EnSCs transfected with Ad-GFP or Ad-shCDC42- GFP following with 72 h of 8Br-cAMPþMPA treatment. (L) Expression of CDC42, P21, and P53 protein levels in primary EnSCs transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (M and N) Representative images of P16 and P21 immunofluorescence staining in primary EnSCs transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (O and P) SA-β-gal staining of primary EnSCs after 48 h of Ad-GFP or Ad-shCDC42-GFP adenovirus treatment and then different days of 8Br-cAMPþMPA treatment. Quantitative analysis of integrated optical density for SA-β-gal staining, with value of the Ad-GFP group at 0 h set as 1. Mean ± SEM. P < 0.05, P < 0.01, P<0.001. ANOVA with Tukey’s multiple comparisons test. Two-way ANOVA with the Bonferroni multiple comparisons test in (P).

Article Snippet: Commercially available ELISA kits were utilized to measure the level of secreted protein (PRL: Elabscience, No. E-EL-H0141; IGFBP1: Elabscience, No. E-EL-H0442; IL6: Elabscience, No. E-EL-H6156; IL8: MULTI SCIENCES, No. EK10896; CLU: Elabscience, No. E-EL-H0038; sST2: Elabscience, No. E-EL-H6082) in collected supernatants according to the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Transfection, Immunofluorescence, Staining

Journal: Translational Vision Science & Technology

Article Title: Proteomic Analysis of Aqueous Humor in Central Retinal Artery Occlusion: Unveiling Novel Insights Into Disease Pathophysiology

doi: 10.1167/tvst.13.8.30

Figure Lengend Snippet: CRAO aqueous humor (AH) proteome. ( A ) Volcano plot of the differentially expressed proteins in CRAO AH as compared to control AH at logFC > |1|, and q value < 0.05. ( B ) Quantification of enolase 2 in plasma samples from controls and patients with CRAO using ELISA. ( C ) Quantification of clusterin in AH samples from controls and patients with CRAO using ELISA. ( D ) Quantification of clusterin in plasma samples from controls and patients with CRAO using ELISA.

Article Snippet: Clusterin, enolase 2, and IGFBP6 were analyzed using human direct sandwich ELISA kits (Protein Tech, cat # KE00110 and KE00050, and Ray Biotech, cat # ELH-IGFBP6-1, respectively) according to manufacturer's instructions.

Techniques: Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Translational Vision Science & Technology

Article Title: Proteomic Analysis of Aqueous Humor in Central Retinal Artery Occlusion: Unveiling Novel Insights Into Disease Pathophysiology

doi: 10.1167/tvst.13.8.30

Figure Lengend Snippet: DR vitreous humor (VH) proteome. ( A ) Volcano plot of the differentially expressed proteins in DR VH as compared to control VH at logFC > |1|, and p value < 0.05. ( B ) Quantification of clusterin in VH samples from controls and patients with DR using ELISA. ( C ) Quantification of enolase 2 in plasma samples from controls and patients with DR using ELISA.

Article Snippet: Clusterin, enolase 2, and IGFBP6 were analyzed using human direct sandwich ELISA kits (Protein Tech, cat # KE00110 and KE00050, and Ray Biotech, cat # ELH-IGFBP6-1, respectively) according to manufacturer's instructions.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Translational Vision Science & Technology

Article Title: Proteomic Analysis of Aqueous Humor in Central Retinal Artery Occlusion: Unveiling Novel Insights Into Disease Pathophysiology

doi: 10.1167/tvst.13.8.30

Figure Lengend Snippet: Shared proteins between the CRAO AH and DR VH datasets. ( A ) Venn diagram of the number of proteins shared between AH and VH proteomic analyses. ( B ) Venn diagram of the number of differentially expressed proteins ( p < 0.05) shared between AH and VH proteomic analyses. ( C ) Quantification of IGFBP6 in plasma samples from controls and patients with CRAO using ELISA. ( D ) Quantification of IGFBP6 in plasma samples from controls and patients with DR using ELISA. ( E ) Heat map of the diseases and functions activated or inhibited in the CRAO AH and DR VH proteomic analyses.

Article Snippet: Clusterin, enolase 2, and IGFBP6 were analyzed using human direct sandwich ELISA kits (Protein Tech, cat # KE00110 and KE00050, and Ray Biotech, cat # ELH-IGFBP6-1, respectively) according to manufacturer's instructions.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

doi: 10.1101/2021.12.15.472789

Figure Lengend Snippet: Schematic overview of the bystander lysis assay. E. coli strains (orange rods) were labelled with convertases (green ovals) in 10% C5-depleted serum and washed. Next, convertase-labelled bacteria (3.3×10 8 per ml) were incubated with: human erythrocytes (1×10 8 per ml), alternative pathway (AP) convertase components (5 nM FB and 20 nM FD) and MAC proteins (100 nM C5, 100 nM C6, 100 nM C7, 100 nM C8 and 500 nM C9). The supernatant was collected after 60 minutes by spinning down bacteria and erythrocytes and analyzed for the presence of haemoglobulin. The percentage of lysed erythrocytes was calculated by setting a buffer only control at 0% lysis and MilliQ control at 100% lysis. b) Bystander erythrocyte lysis for MAC-sensitive (MAC-sens) and MAC-resistant (MAC-res) E. coli strains. c) Bystander erythrocyte lysis for convertase-labelled MAC-resistant E. coli 552059.1 incubated with 10% pooled human serum, MAC proteins (30 nM C5, 30 nM C6, 30 nM C7, 30 nM C8 and 300 nM C9) or MAC components with 133 nM vitronectin (Vn), 133 nM clusterin (Clu) or 20 µg/ml C5 conversion inhibitor OmCI. Data represent individual values with mean +/- SD of three independent experiments. Statistical analysis was done using an ordinary one-way ANOVA with Tukey’s multiple comparisons test (c) and relevant p-values are indicated in the figure (all conditions compared with MAC proteins only).

Article Snippet: Vitronectin (plasma isolated) was obtained from Advanced Biomatrix and recombinantly expressed human clusterin from R&D systems.

Techniques: Lysis, Bacteria, Incubation, Control

Journal: Scientific Reports

Article Title: Clusterin mediates hydroquinone-induced cytotoxic responses in HL-60 differentiated cells

doi: 10.1038/s41598-024-82140-0

Figure Lengend Snippet: CLU protein concentration in HL-60 cells exposed to different concentrations of HQ. CLU protein levels were quantified using ELISA after treatment with HQ at concentrations of 0, 10, 25, and 50 µmol/L ( n = 3 per group). The error bars represent SD, and the mean is shown as horizontal bars. The control group (0 µmol/L HQ) exhibited the highest CLU concentration (19.05 ± 2.32 ng/mL), while HQ-treated groups showed a progressive decrease: 14.45 ± 1.23 ng/mL at 10 µmol/L, 9.43 ± 0.40 ng/mL at 25 µmol/L, and 9.46 ± 0.67 ng/mL at 50 µmol/L. Significant differences were determined by ANOVA followed by Tukey post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05), ns : not significant.

Article Snippet: The concentration of sCLU proteins was measured using the Human Clusterin ELISA Kit (Boster Biological Technology, China) according to the instructions.

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay, Control, Concentration Assay

Journal: Scientific Reports

Article Title: Clusterin mediates hydroquinone-induced cytotoxic responses in HL-60 differentiated cells

doi: 10.1038/s41598-024-82140-0

Figure Lengend Snippet: Concentration of sCLU protein in the cell culture supernatant after exposure to varying HQ concentrations. The concentration of sCLU protein in the supernatant of HL -60 cell cultures was determined using ELISA following treatment with HQ at 0, 10, 25, and 50 µmol/L ( n = 3 per group). The control group (0 µmol/L HQ) exhibited the highest concentration (25.88 ± 4.24 ng/mL), while the 10 µmol/L HQ group showed a slight, non-significant decrease (23.73 ± 2.03 ng/mL, p = 0.696). Significant reductions were observed in the 25 µmol/L (17.45 ± 0.36 ng/mL) and 50 µmol/L (12.96 ± 0.67 ng/mL) groups compared to the control (* p < 0.05, *** p < 0.001, ns p > 0.05), ns : not significant.

Article Snippet: The concentration of sCLU proteins was measured using the Human Clusterin ELISA Kit (Boster Biological Technology, China) according to the instructions.

Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Control