Human CCL2/JE/MCP-1 Antibody

Detection of Human CCL2/JE/MCP-1 by Immunohistochemistry

Expression patterns of CCL2, CCL5, TNF alpha and IL-1 beta in healthy individuals and breast cancer patients. Representative examples of the expression of CCL2, CCL5, TNF alpha and IL-1 beta in the different groups of patients included in the study, in biopsies obtained at the time of diagnosis. (a1-a4) Patients diagnosed with benign breast disorders. The pictures demonstrate the lack of staining of the four factors in the normal breast epithelial cells, as denoted in the majority of patients included in this group. (b1-b4) DCIS patients. The pictures demonstrate positive staining of the four factors in the malignant lesions, as denoted in the majority of patients included in this group. (c1-c4) IDC-no-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (d1-d4) IDC-with-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (a1, b1, c1, d1) CCL2 staining; (a2, b2, c2, d2) CCL5 staining; (a3, b3, c3, d3) TNF alpha staining; (a4, b4, c4, d4) IL-1 beta staining. The expression of the proteins was determined by IHC using specific antibodies, whose specificity in IHC was verified. The values of photo magnification are indicated in the left bottom corner of each of the pictures. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/21486440), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by Immunohistochemistry

Expression patterns of CCL2, CCL5, TNF alpha and IL-1 beta in healthy individuals and breast cancer patients. Representative examples of the expression of CCL2, CCL5, TNF alpha and IL-1 beta in the different groups of patients included in the study, in biopsies obtained at the time of diagnosis. (a1-a4) Patients diagnosed with benign breast disorders. The pictures demonstrate the lack of staining of the four factors in the normal breast epithelial cells, as denoted in the majority of patients included in this group. (b1-b4) DCIS patients. The pictures demonstrate positive staining of the four factors in the malignant lesions, as denoted in the majority of patients included in this group. (c1-c4) IDC-no-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (d1-d4) IDC-with-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (a1, b1, c1, d1) CCL2 staining; (a2, b2, c2, d2) CCL5 staining; (a3, b3, c3, d3) TNF alpha staining; (a4, b4, c4, d4) IL-1 beta staining. The expression of the proteins was determined by IHC using specific antibodies, whose specificity in IHC was verified. The values of photo magnification are indicated in the left bottom corner of each of the pictures. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/21486440), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by Immunohistochemistry

Expression patterns of CCL2, CCL5, TNF alpha and IL-1 beta in healthy individuals and breast cancer patients. Representative examples of the expression of CCL2, CCL5, TNF alpha and IL-1 beta in the different groups of patients included in the study, in biopsies obtained at the time of diagnosis. (a1-a4) Patients diagnosed with benign breast disorders. The pictures demonstrate the lack of staining of the four factors in the normal breast epithelial cells, as denoted in the majority of patients included in this group. (b1-b4) DCIS patients. The pictures demonstrate positive staining of the four factors in the malignant lesions, as denoted in the majority of patients included in this group. (c1-c4) IDC-no-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (d1-d4) IDC-with-relapse patients. The pictures demonstrate positive staining of the four factors in the tumor cells, as denoted in the majority of patients included in this group. (a1, b1, c1, d1) CCL2 staining; (a2, b2, c2, d2) CCL5 staining; (a3, b3, c3, d3) TNF alpha staining; (a4, b4, c4, d4) IL-1 beta staining. The expression of the proteins was determined by IHC using specific antibodies, whose specificity in IHC was verified. The values of photo magnification are indicated in the left bottom corner of each of the pictures. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/21486440), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by ELISA

Induction of CCL2 and CLXL8 in TNF alpha-stimulated MSCs is not mediated via the AP-1 pathway. (A) Human BM-derived MSCs were stimulated by TNF-alpha (50 ng/ml) for 5 and 10 minutes. Control cells were treated by the vehicle of TNF-alpha. c-Jun levels and phosphorylation were determined by western blot (WB) analyses. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. (B) Human BM-derived MSCs were transiently transfected by small interfering RNA (siRNA) to c-Jun or by control siRNA. (B1) c-Jun expression was determined by WB analyses. beta-Tubulin was used as loading control. (B2) Following siRNA transfection, the cells were stimulated by TNF-alpha (25 ng/ml; in this part of the study we used a suboptimal concentration of TNF-alpha in order to facilitate detection of inhibitory effects) for 24 hours. Expression levels of CCL2 and CXCL8 in the supernatants of the cells were determined by ELISA, in the linear range of absorbance. #siRNA to c-Jun has yielded minor increases or reductions in CCL2 and CXCL8 secretion in different experiments (see Results and discussion), and thus overall there was no significant effect on CCL2 and CXCL8 secretion. In all panels, the findings are representatives of n = 3 independent experiments that have shown similar results. Image collected and cropped by CiteAb from the following publication (https://stemcellres.biomedcentral.com/articles/10.1186/s13287-015-0080-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by Western Blot

NF-kappa B is essential in mediating TNF-alpha -induced release of chemokines by MSCs. (A) Human BM-derived MSCs were stimulated by TNF-alpha (50 ng/ml) for 15 minutes. The levels of I kappaB alpha (the negative regulator of the NF-kappa B pathway) were determined by WB analyses. GAPDH was used as a loading control throughout. (B) CAFs were generated by culturing MSCs with Tumor CM from MDA-MB-231 (MDA) or MCF-7 breast tumor cells over a prolonged period of time (~30 days). TNF-alpha (50 ng/ml) was added for the last 24 hours to MSCs + Tumor CM cells and I kappaB alpha levels were determined by WB analyses. (C) CAF #1 cells were stimulated for 48 hours by TNF-alpha (50 ng/ml). I kappaB alpha levels were determined by WB analyses. (D) Human BM-derived MSCs were stimulated with TNF-alpha (50 ng/ml) for 10 minutes. p65 phosphorylation was determined by WB analyses. (E) Human BM-derived MSCs were transiently transfected by siRNA to p65 or by control siRNA. (E1) p65 expression was determined by WB analyses. (E2) Following siRNA transfection, the cells were stimulated by TNF-alpha (25 ng/ml; a suboptimal concentration of TNF-alpha in order to facilitate detection of inhibitory effects) for 48 hours. Expression of CCL2 and CXCL8 in the supernatants of the cells was determined by ELISA, in the linear range of absorbance. In all panels, the findings are representatives of n = 3 independent experiments that have shown similar results. Image collected and cropped by CiteAb from the following publication (https://stemcellres.biomedcentral.com/articles/10.1186/s13287-015-0080-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by ELISA

Microparticle composition influences inflammatory cytokine production from activated monocytes.THP-1 cells (A) or CD14 sorted blood monocytes (B) were treated with control microparticles (grey bars) or PPAR gamma-overexpressing microparticles (black bars) or no microparticles (white bars) for 4 hours before activation with LPS or PAM3CSK4 for 24 hours. Supernatants were collected and pro-inflammatory cytokines IL-8 (top), MCP-1 (bottom) were measured by ELISA. Dotted line represents baseline cytokine production from unactivated cells with no microparticle exposure. Mean values with standard errors represent one of at least 3 experiments. Two-way ANOVA with Tukey's multiple comparison post test was performed to determine statistical significance. * indicates (p<0.05). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25426628), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CCL2/JE/MCP-1 by ELISA

Microparticle composition influences inflammatory cytokine production from activated monocytes.THP-1 cells (A) or CD14 sorted blood monocytes (B) were treated with control microparticles (grey bars) or PPAR gamma-overexpressing microparticles (black bars) or no microparticles (white bars) for 4 hours before activation with LPS or PAM3CSK4 for 24 hours. Supernatants were collected and pro-inflammatory cytokines IL-8 (top), MCP-1 (bottom) were measured by ELISA. Dotted line represents baseline cytokine production from unactivated cells with no microparticle exposure. Mean values with standard errors represent one of at least 3 experiments. Two-way ANOVA with Tukey's multiple comparison post test was performed to determine statistical significance. * indicates (p<0.05). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25426628), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CCL2/JE/MCP-1 in human Crohn's disease.

Formalin-fixed paraffin-embedded tissue sections of human Crohn’s Disease were probed for CCL2 mRNA (ACD RNAScope Probe, catalog # 423811; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using mouse anti-human CCL2 monoclonal antibody (R&D Systems catalog # MAB679) at 20ug/mL with 1 hour incubation at room temperature followed by incubation with anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and membrane.