human ccl2 elisa kit  (R&D Systems)

Journal: Renal Failure

Article Title: Assessment of urine CCL2 as a potential diagnostic biomarker for acute kidney injury and septic acute kidney injury in intensive care unit patients

doi: 10.1080/0886022X.2024.2313171

Figure Lengend Snippet: Patients’ clinical characteristics.

Article Snippet: The CCL2 levels in plasma and urine were measured using a commercial human CCL2 ELISA kit (R&D Systems, Bio-Techne, Minneapolis, USA) following the manufacturer’s instructions.

Techniques: Control

Journal: Renal Failure

Article Title: Assessment of urine CCL2 as a potential diagnostic biomarker for acute kidney injury and septic acute kidney injury in intensive care unit patients

doi: 10.1080/0886022X.2024.2313171

Figure Lengend Snippet: Plasma and urine CCL2 levels between AKI and non-AKI patients at hospital and ICU admission. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The CCL2 levels in plasma and urine were measured using a commercial human CCL2 ELISA kit (R&D Systems, Bio-Techne, Minneapolis, USA) following the manufacturer’s instructions.

Techniques: Clinical Proteomics

Journal: Renal Failure

Article Title: Assessment of urine CCL2 as a potential diagnostic biomarker for acute kidney injury and septic acute kidney injury in intensive care unit patients

doi: 10.1080/0886022X.2024.2313171

Figure Lengend Snippet: The predictive performance of urine CCL2 for AKI by ROC analysis. (A) ROC curve. (B) AUC and prediction sensitivity and specificity.

Article Snippet: The CCL2 levels in plasma and urine were measured using a commercial human CCL2 ELISA kit (R&D Systems, Bio-Techne, Minneapolis, USA) following the manufacturer’s instructions.

Techniques:

Journal: Renal Failure

Article Title: Assessment of urine CCL2 as a potential diagnostic biomarker for acute kidney injury and septic acute kidney injury in intensive care unit patients

doi: 10.1080/0886022X.2024.2313171

Figure Lengend Snippet: Plasma and urine CCL2 levels between SAKI and non-septic AKI patients at hospital and ICU admission. *** p < 0.001.

Article Snippet: The CCL2 levels in plasma and urine were measured using a commercial human CCL2 ELISA kit (R&D Systems, Bio-Techne, Minneapolis, USA) following the manufacturer’s instructions.

Techniques: Clinical Proteomics

Journal: Renal Failure

Article Title: Assessment of urine CCL2 as a potential diagnostic biomarker for acute kidney injury and septic acute kidney injury in intensive care unit patients

doi: 10.1080/0886022X.2024.2313171

Figure Lengend Snippet: The predictive performance of urine CCL2 for SAKI by ROC analysis. (A) ROC curve. (B) AUC and prediction sensitivity and specificity.

Article Snippet: The CCL2 levels in plasma and urine were measured using a commercial human CCL2 ELISA kit (R&D Systems, Bio-Techne, Minneapolis, USA) following the manufacturer’s instructions.

Techniques:

Journal: British journal of cancer

Article Title: Chromatin accessibility uncovers KRAS-driven FOSL2 promoting pancreatic ductal adenocarcinoma progression through up-regulation of CCL28.

doi: 10.1038/s41416-023-02313-y

Figure Lengend Snippet: Fig. 7 FOSL2 promotes Treg cell recruitment through CCL28. a–d Scheme representing the experimental procedure (a), representative tumour image (b), tumour weight (c), and tumour growth curves (d) of C57BL/6 mice injected subcutaneously with Vector and oe-FOSL2 PanO2 cells with treatment of CCL28 neutralising antibody or IgG. e Representative flow staining (left panel) and frequency (right panel) of Treg cells in tumour xenograft of indicated groups. f Representative flow staining (left panel) and frequency (right panel) of CD8+ T cells in tumour xenograft of indicated groups. g The frequency of CD4+ T cells in tumour xenograft of indicated groups. h The level of VEGFA protein in the serum of Vector or oe-FOSL2 mice following administration of anti-CCL28 antibody or IgG, as determined by ELISA. i Representative immunofluorescence staining (left panel) and quantitation (right panel) of mean fluorescence intensity of CD31 in indicated groups. Data are representative shown as mean ± SD or SEM (**P < 0.01; ***P < 0.001; ns not significant).

Article Snippet: The concentrations of CCL28 and VEGFA in the culture supernatant of human PDAC cell lines or mouse serum were measured using the CCL28 ELISA kit (human, DY717; mouse, DY533) or VEGFA ELISA kit (mouse, DY493) following the manufacturer’s instructions (R&D Systems).

Techniques: Injection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay, Quantitation Assay

Journal: Journal of reproductive immunology

Article Title: Detection of CCL25 and the correlation between CCL25, CCL28, IL-7, and TSLP in human breast milk.

doi: 10.1016/j.jri.2022.103783

Figure Lengend Snippet: Fig. 1. Comparison of chemokine and cytokine levels between colostrum and mature milk. The concentrations of chemokines and cytokines in colostrum and mature milk were analyzed: (a) CCL25, (b) CCL28, (c) IL-7, and (d) TSLP. The levels of each of the chemokines and cytokines were compared between colostrum and mature milk using Wilcoxon signed rank test. Medians of chemokine and cytokine levels in BM are shown with a bar. An asterisk indicates a significant difference at P < 0.05.

Article Snippet: CCL25, CCL28, IL-7, and TSLP levels in whey were measured using Human CCL25 DuoSet ELISA kit (R&D Systems), Human CCL28 DuoSet ELISA kit (R&D Systems), Human IL-7 DuoSet ELISA kit (R&D Systems), and TSLP Human Uncoated ELISA Kit (Invitrogen) according with the previous reports (Aihara et al., 2014; Hossny et al., 2020; Pallister et al., 2015).

Techniques: Comparison

Journal: Journal of reproductive immunology

Article Title: Detection of CCL25 and the correlation between CCL25, CCL28, IL-7, and TSLP in human breast milk.

doi: 10.1016/j.jri.2022.103783

Figure Lengend Snippet: Fig. 2. Comparison of chemokine and cytokine levels in BM between primiparous and multiparous women.CCL25, CCL28, TSLP, and IL-7 levels in BM were compared between primiparous and multiparous women: (a) CCL25, (b) CCL28, (c) TSLP, and (d) IL-7. Medians of chemokine and cytokine levels in BM are shown with a bar. Primi. and Multi. indicate primiparous and multiparous women, respectively. Cytokine levels in colostrum and mature milk are shown with triangles and circles, respectively.

Article Snippet: CCL25, CCL28, IL-7, and TSLP levels in whey were measured using Human CCL25 DuoSet ELISA kit (R&D Systems), Human CCL28 DuoSet ELISA kit (R&D Systems), Human IL-7 DuoSet ELISA kit (R&D Systems), and TSLP Human Uncoated ELISA Kit (Invitrogen) according with the previous reports (Aihara et al., 2014; Hossny et al., 2020; Pallister et al., 2015).

Techniques: Comparison

Journal: Journal of reproductive immunology

Article Title: Detection of CCL25 and the correlation between CCL25, CCL28, IL-7, and TSLP in human breast milk.

doi: 10.1016/j.jri.2022.103783

Figure Lengend Snippet: Fig. 1. Comparison of chemokine and cytokine levels between colostrum and mature milk. The concentrations of chemokines and cytokines in colostrum and mature milk were analyzed: (a) CCL25, (b) CCL28, (c) IL-7, and (d) TSLP. The levels of each of the chemokines and cytokines were compared between colostrum and mature milk using Wilcoxon signed rank test. Medians of chemokine and cytokine levels in BM are shown with a bar. An asterisk indicates a significant difference at P < 0.05.

Article Snippet: CCL25, CCL28, IL-7, and TSLP levels in whey were measured using Human CCL25 DuoSet ELISA kit (R&D Systems), Human CCL28 DuoSet ELISA kit (R&D Systems), Human IL-7 DuoSet ELISA kit (R&D Systems), and TSLP Human Uncoated ELISA Kit (Invitrogen) according with the previous reports (Aihara et al., 2014; Hossny et al., 2020; Pallister et al., 2015).

Techniques: Comparison

Journal: Journal of reproductive immunology

Article Title: Detection of CCL25 and the correlation between CCL25, CCL28, IL-7, and TSLP in human breast milk.

doi: 10.1016/j.jri.2022.103783

Figure Lengend Snippet: Fig. 2. Comparison of chemokine and cytokine levels in BM between primiparous and multiparous women.CCL25, CCL28, TSLP, and IL-7 levels in BM were compared between primiparous and multiparous women: (a) CCL25, (b) CCL28, (c) TSLP, and (d) IL-7. Medians of chemokine and cytokine levels in BM are shown with a bar. Primi. and Multi. indicate primiparous and multiparous women, respectively. Cytokine levels in colostrum and mature milk are shown with triangles and circles, respectively.

Article Snippet: CCL25, CCL28, IL-7, and TSLP levels in whey were measured using Human CCL25 DuoSet ELISA kit (R&D Systems), Human CCL28 DuoSet ELISA kit (R&D Systems), Human IL-7 DuoSet ELISA kit (R&D Systems), and TSLP Human Uncoated ELISA Kit (Invitrogen) according with the previous reports (Aihara et al., 2014; Hossny et al., 2020; Pallister et al., 2015).

Techniques: Comparison

Journal: Journal of Inflammation Research

Article Title: IFN-γ and LPS Induce Synergistic Expression of CCL2 in Monocytic Cells via H3K27 Acetylation

doi: 10.2147/jir.s368352

Figure Lengend Snippet: Figure 1 IFN-γ and LPS cooperatively induce CCL2 in monocytic cells/macrophages. THP-1 monocytic cells, primary monocytes, and THP-1-derived macrophages were treated for 24h with IFN-γ (10 ng/mL) alone or in combination with LPS (10 ng/mL). Total RNA was extracted and Ccl2 mRNA expression was quantified by real-time RT-PCR. CCL2 protein was measured in cell supernatants using commercial ELISA kit. All data are expressed as mean ± SEM (n ≥ 3). Group means between two data sets were compared using Student’s t-test and those of more than two data sets were compared using one-way ANOVA with post-hoc Tukey’s test. All p-values < 0.05 were considered significant (ns, non- significant, *p< 0.05, **p< 0.01, and ***p< 0.001). Elevated CCL2 gene (A) and secreted protein (B) expression is shown in THP-1 monocytic cells co-stimulated with IFN-γ and LPS compared to cells treated with IFN-γ and LPS alone. Primary monocytes (C) also show increased CCL2 secreted protein expression after co-stimulation with IFN-γ and LPS compared to stimulation with either IFN-γ or LPS. In addition to monocytic cells and primary monocytes, THP-1-derived macrophages co-stimulated with IFN-γ and LPS also display elevated expression of CCL2 transcripts (D) and secreted protein (E). (F and G) THP-1 cells were primed with IFN-γ and LPS, separately, followed by LPS and IFN-γ treatment, respectively, for 24h. Ccl2 mRNA and protein expression was quantified and statistically analyzed as described above. The data show that only the IFN-γ priming followed by LPS stimulation led to elevated CCL2 mRNA (F) and secreted protein (G) expression in monocytic cells compared to controls stimulated with IFN-γ or LPS alone.

Article Snippet: CCL2-secreted protein levels were measured in the supernatants of THP-1 cells stimulated with IFN-γ (10 ng/mL), LPS (10 ng/mL), alone or in combination, using human CCL2 DuoSet ELISA kit and following the manufacturer’s instructions (Cat. # DY 279, R&D Systems Inc.).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: IFN-γ and LPS Induce Synergistic Expression of CCL2 in Monocytic Cells via H3K27 Acetylation

doi: 10.2147/jir.s368352

Figure Lengend Snippet: Figure 2 Synergistic expression of CCL2 is dependent on STAT1. (A) THP-1 monocytic cells were transfected with control/scrambled siRNA or STAT1 siRNA and incubated for 40h. Total RNA was extracted and real-time RT-PCR was performed to measure STAT1 mRNA expression. Target mRNA levels were normalized against GAPDH mRNA and gene expression relative to control was calculated using 2−ΔΔCT method. Relative STAT1 mRNA expression was expressed as fold expression over average of control (scrambled siRNA) gene expression. All data are expressed as mean ± SEM (n ≥ 3) and group means between two data sets were compared using Student’s t-test. All p-values < 0.05 were considered significant. The data show significant suppression of STAT1 mRNA expression in cells transfected with STAT1 siRNA compared to control siRNA transfected cells (**p< 0.01). (B and C) STAT1-deficient cells were treated for 24h with IFN-γ (10 ng/mL) and/or LPS (10 ng/mL) and Ccl2 mRNA (B) and protein (C) expression was determined using real-time RT-PCR and ELISA, respectively. Target mRNA levels were normalized against GAPDH mRNA and gene expression relative to control was calculated using 2−ΔΔCT method. Relative CCL2 mRNA expression was expressed as fold change over average of control (vehicle treatment) gene expression. All data are expressed as mean ± SEM (n ≥ 3) and group means between two data sets were compared using Student’s t-test. The data show significant suppression of (B) CCL2 mRNA (***p< 0.001) and (C) CCL2 secreted protein (**p< 0.01) in cells co-stimulated with IFN-γ and LPS as compared to those stimulated with IFN-γ or LPS alone. (D) Western blot showing phosphorylation of STAT1 after IFN-γ (10 ng/mL) treatment over time indicates the optimal STAT1 phosphorylation at 120 min.

Article Snippet: CCL2-secreted protein levels were measured in the supernatants of THP-1 cells stimulated with IFN-γ (10 ng/mL), LPS (10 ng/mL), alone or in combination, using human CCL2 DuoSet ELISA kit and following the manufacturer’s instructions (Cat. # DY 279, R&D Systems Inc.).

Techniques: Expressing, Transfection, Control, Incubation, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay, Western Blot, Phospho-proteomics

Journal: Journal of Inflammation Research

Article Title: IFN-γ and LPS Induce Synergistic Expression of CCL2 in Monocytic Cells via H3K27 Acetylation

doi: 10.2147/jir.s368352

Figure Lengend Snippet: Figure 3 H3K27 acetylation levels at different sites of CCL2 promotor region. (A) THP-1 monocytic cells were treated with IFN-γ (10 ng/mL) and/or LPS (10 ng/mL) for 4 h and the treatment with vehicle alone served as control. Cell lysates were used for determination of H3K27 acetylation by Western blotting. Briefly, cell lysates were resolved using 12% SDS-PAGE and blots were probed with rabbit anti-human H3K27 antibody (1:1000 dilution) at 4°C overnight. Blots were washed, incubated for 2h with HRP-conjugated secondary antibody (1:2500 dilution), and immunoreactive bands were developed and visualized using ChemiDoc™ MP Imaging Systems. (B) Western blot band densities were quantified and data were expressed as mean ± SEM (n = 3) values which were compared for various treatments using one-way ANOVA and post-hoc Tukey’s test. All p-values < 0.05 were considered significant (ns, non-significant, *p< 0.05, **p< 0.01, and ***p< 0.001). The data show increased H3K27 acetylation (H3K27 ac) in the cells stimulated with IFN-γ or IFN-γ+LPS compared to cells stimulated with LPS only (***p< 0.001). (C) The schematic diagram of CCL2 gene promotor region is shown. (D–I) THP1 cells were stimulated with IFN-γ (10 ng/mL) and/or LPS (10 ng/mL) by incubation for 24h at 37°C. Chromatin immunoprecipitation was done on cell lysate as described in methodology using antibodies specific to H3K27, Histone H3, and normal rabbit IgG for overnight at 4° C. H3K27 acetylation levels induced by treatments compared to control (vehicle) were detected by qPCR using primers specific to the closest regions of transcription start site at the CCL2 promotor. Data (mean ± SEM, n = 3) were expressed as fold enrichment levels and were compared for different treatments against control using one-way ANOVA with post-hoc Tukey’s test. All p-values < 0.05 were considered significant (*p< 0.05 and **p< 0.01).

Article Snippet: CCL2-secreted protein levels were measured in the supernatants of THP-1 cells stimulated with IFN-γ (10 ng/mL), LPS (10 ng/mL), alone or in combination, using human CCL2 DuoSet ELISA kit and following the manufacturer’s instructions (Cat. # DY 279, R&D Systems Inc.).

Techniques: Control, Western Blot, SDS Page, Incubation, Imaging, Chromatin Immunoprecipitation

Journal: Journal of Inflammation Research

Article Title: IFN-γ and LPS Induce Synergistic Expression of CCL2 in Monocytic Cells via H3K27 Acetylation

doi: 10.2147/jir.s368352

Figure Lengend Snippet: Figure 4 Histone acetyltransferases (HATs) and histone deacetylases (HDACs) regulate synergy between IFN-γ and LPS for the CCL2 production. THP-1 monocytic cells were treated with anacardic acid (HATs inhibitor; 50 μM) overnight or with TSA (HDACs inhibitor; 25 nM) for 6 h, followed by treatments with IFN-γ (10 ng/mL) and/or LPS (10 ng/mL) for 24h, and the treatment with vehicle alone served as control. Ccl2 mRNA expression was determined by real-time RT-PCR and target gene expression were normalized to GAPDH expression. Relative changes in Ccl2 gene expression were calculated using 2−ΔΔCT method and expressed as fold change over its expression in control (vehicle treatment). CCL2 secreted protein expression was detected by ELISA as described in materials and methods. All data were expressed as mean ± SEM values (n = 3) and group means between two data sets were compared using Student’s t-test. All p-values < 0.05 were considered significant (**p<0.01, ***p<0.001). The data show the reduced expression of CCL2 (A) mRNA (***p<0.001) and (B) secreted protein (**p<0.01) in the cells that were treated with HAT-inhibitor anacardic acid before co-stimulation with IFN-γ and LPS as compared to similarly stimulated cells that were not pre-treated with anacardic acid. Interestingly, increased expression of CCL2 (C) mRNA (**p<0.01) and (D) secreted protein (***p<0.001) were observed in the cells that were treated with HDAC-inhibitor TSA before stimulation with LPS only as compared to similarly stimulated cells that were not pre-treated with TSA, suggesting that TSA priming could mimic effect of and substitute for IFN-γ in cooperativity with LPS.

Article Snippet: CCL2-secreted protein levels were measured in the supernatants of THP-1 cells stimulated with IFN-γ (10 ng/mL), LPS (10 ng/mL), alone or in combination, using human CCL2 DuoSet ELISA kit and following the manufacturer’s instructions (Cat. # DY 279, R&D Systems Inc.).

Techniques: Control, Expressing, Quantitative RT-PCR, Targeted Gene Expression, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: mSphere

Article Title: CCL19 and CCL28 Assist Herpes Simplex Virus 2 Glycoprotein D To Induce Protective Systemic Immunity against Genital Viral Challenge

doi: 10.1128/mSphere.00058-21

Figure Lengend Snippet: gD-specific IgG and IgA responses and Ig subclasses in serum postchallenge. Serum gD-specific IgG (A) and IgA (B) at indicated days postboost and postchallenge. The endpoint titrations of gD-specific IgG and IgA were determined by ELISA separately. (C) The distribution of gD-specific IgG1, IgG2a, IgG2b, IgG3, and IgM isotypes in murine sera at 2 weeks postboost (upper) and 9 dpi (lower). (D) gD-specific IgG2a/IgG1 ratio of the different groups at 2 weeks postboost and 9 dpi. The quantification of each Ig subclass was measured by ELISA and calculated according to the standard curve obtained using corresponding Ig subclass standards. CCL19, 20 mol; CCL28, 10 mol. Data are the means ± SEM, n = 5 or 7 mice per group, from at least two independent experiments with each condition being performed in duplicate. Statistically significant differences determined by comparison to the pgD + pcDNA3.1 group are indicated. NS, not statistically significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Chemokines were detected using the CCL19 and CCL28 DuoSet ELISA kit (R&D Systems, USA), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: mSphere

Article Title: CCL19 and CCL28 Assist Herpes Simplex Virus 2 Glycoprotein D To Induce Protective Systemic Immunity against Genital Viral Challenge

doi: 10.1128/mSphere.00058-21

Figure Lengend Snippet: Ag-specific Th1/Th2-associated cytokine production from splenocytes and antisera. (A) Cytokine production in splenocyte supernatants from immunized mice (2 weeks postboost) and antisera after HSV-2 challenge (5 dpi and 9 dpi) was quantified by flow cytometry. (B) Fold changes in the pgD + pCCL28 or pCCL28-IZ-gD groups over the pgD + pCCL19 group. Splenocytes (1 × 10 7 cells) isolated from all groups postboost were stimulated with purified gD (2 μg/ml) and cocultivated for 5 days, and then the supernatants were collected. (C) Hot map of cytokine fold changes (i.e., pgD + CCL28/pgD + CCL19 and pCCL28-IZ-gD/pgD + CCL19). Group 1, 1-1, and 1-2 is named pgD + CCL28/pgD + CCL19, and 2, 2-1, and 2-2 is named pCCL28-IZ-gD/pgD + CCL19. Antisera were collected at 5 and 9 dpi. The production of Th1 (IL-2, IFN-γ, and TNF-α)- and Th2 (IL-4 and IL-5)-associated cytokines was detected using a CBA kit. Data shown are representative of the mean cytokine concentrations (pg/ml) ± SEM with n = 5 mice per group. Statistically significant differences determined by comparing to the pgD + pcDNA3.1 group are indicated. NS, not statistically significant; *, P < 0.05; ***, P < 0.001.

Article Snippet: Chemokines were detected using the CCL19 and CCL28 DuoSet ELISA kit (R&D Systems, USA), respectively.

Techniques: Flow Cytometry, Isolation, Purification

Journal: mSphere

Article Title: CCL19 and CCL28 Assist Herpes Simplex Virus 2 Glycoprotein D To Induce Protective Systemic Immunity against Genital Viral Challenge

doi: 10.1128/mSphere.00058-21

Figure Lengend Snippet: Responsive immunocyte migration to secondary lymph nodes postimmunization and postchallenge. IgA + cells at colorectal sites mediated by the CCL19 or CCL28 adjuvant in immunized mice. (A) IgA + cells in murine colorectal mucosal samples were stained with DBA and hematoxylin. Data shown are representative immunohistochemistry results (magnification, ×200). (B) Mean counts of IgA + cells of 10 high-power fields for each group. Migration of murine splenocytes and MLNLs for each group in response to murine CCL19 or CCL28 protein. Single lymphocytes were prepared and counted for the chemotactic response to CCL19 or CCL28 using a Transwell system, and the fold changes were calculated compared to the cell number in the lower chamber without CCL19 (C) or CCL28 (D). The frequencies of CCR7 +/− (E) and CCR3 + (F) CD3 + splenocytes at 2 weeks postboost and 9 dpi were analyzed by flow cytometry. By comparing to the pgD + pcDNA3.1 group, the fold change of CCR7 − (G) and CCR3 + (H) CD3 + splenocyte frequencies in the pCCL19 or pCCL28 adjuvant groups were calculated, and the gating strategies are shown in . Data shown are the means ± SEM for each group ( n = 5 mice/group). Statistically significant differences determined by comparing to the pgD + pcDNA3.1 group are indicated. NS, not statistically significant; ***, P < 0.001.

Article Snippet: Chemokines were detected using the CCL19 and CCL28 DuoSet ELISA kit (R&D Systems, USA), respectively.

Techniques: Migration, Adjuvant, Staining, Immunohistochemistry, Flow Cytometry

Journal: American Journal of Clinical and Experimental Immunology

Article Title: The CCL28 levels are elevated in the serum of patients with irritable bowel syndrome and associated with the clinical symptoms

doi:

Figure Lengend Snippet: The levels of CCL28 is higher in patients than in the control group. The levels of CCL28 was detected in the serum of 41 patients and 41 normal individuals by Elisa. The average of CCL28 in patients and normal individuals were 293±113 and 228±114, respectively.

Article Snippet: The concentration of CCL28 levels were examined using solid-phase sandwich ELISA kits according to the manufacturer’s instructions (Cat# DY717, R & D Systems, San Diego, CA).

Techniques: Control, Enzyme-linked Immunosorbent Assay