c5ar monoclonal anti human antibodies  (Hycult Biotech)

Journal: Journal of Clinical Medicine

Article Title: Role of C5aR1 and C5L2 Receptors in Ischemia-Reperfusion Injury

doi: 10.3390/jcm10050974

Figure Lengend Snippet: Gene expression of C5aR1 , C5L2 , complement regulators, an inflammatory and profibrotic gene from in vitro ischemia-reperfusion injury (IRI) model. Quantitative real-time RT–PCR for analyzing C5aR1 , C5L2 , CD59 , KIM-1 , Colagen1A1 , and Vimentin was performed in HK-2 cells collected after a reoxygenation step from the IRI model. Gene expression was expressed as relative gene expression vs. PPIA . Data were shown as mean ± SEM and analyzed by the Mann–Whitney test. * p < 0.05 vs. normoxia + 10% FBS condition. $ p < 0.05 vs. N+NHS, # p < 0.05 vs. H+FBS.

Article Snippet: After drying at room temperature, we fixed the samples in 10% buffered formalin for 10 min. After PBS washing, 3% goat serum in PBS was used to block nonspecific binding sites for 1 h. Primary antibodies against C5aR1 (#HM2094, Hycult Biotech) (mAb 1:10) and C5aL (#HP9036, Hycult Biotech) (pAb 1:25) were then incubated for one hour more.

Techniques: Expressing, In Vitro, Quantitative RT-PCR, MANN-WHITNEY

Journal: Journal of Clinical Medicine

Article Title: Role of C5aR1 and C5L2 Receptors in Ischemia-Reperfusion Injury

doi: 10.3390/jcm10050974

Figure Lengend Snippet: Cellular distribution of C5aR1. HK-2 under normoxic or hypoxic conditions for 48 h and later reoxygenated for 30 min with 10% heat-inactivated fetal bovine serum (FBS) or 10% normal human serum (NHS). Double immunostaining was performed with C5aR1 (green) and β-actin (red). Nuclei were visualized by DAPI (blue) (scale bar: 50 μm). The intensity of the staining was expressed as arbitrary units of fluorescence (AUF) per cell. Data from 3 different experiments are shown as mean ± SEM and analyzed by the Mann–Whitney test. The experimental conditions were: normoxia + 10% FBS (N+FBS), normoxia + 10% NHS (N+NHS), hypoxia + 10% FBS (H+FBS) and hypoxia + 10% NHS (H+NHS). * p < 0.05 vs. normoxia + 10% FBS condition.

Article Snippet: After drying at room temperature, we fixed the samples in 10% buffered formalin for 10 min. After PBS washing, 3% goat serum in PBS was used to block nonspecific binding sites for 1 h. Primary antibodies against C5aR1 (#HM2094, Hycult Biotech) (mAb 1:10) and C5aL (#HP9036, Hycult Biotech) (pAb 1:25) were then incubated for one hour more.

Techniques: Double Immunostaining, Staining, Fluorescence, MANN-WHITNEY

Journal: Journal of Clinical Medicine

Article Title: Role of C5aR1 and C5L2 Receptors in Ischemia-Reperfusion Injury

doi: 10.3390/jcm10050974

Figure Lengend Snippet: C5aR1 and C5L2 stain in biopsies from kidney transplantation (KT) patients with DGF and one-year protocol biopsies without tissue damage as a control group. C5aR1 and C5l2 were evaluated in the tubular compartment using immunohistochemistry and semiquantitative scoring. C5aR1 stain was observed in the apical and basal side of the membrane from tubular epithelial cells, while C5L2 deposit was observed in the endothelium from peritubular capillaries. DGF biopsies showed more frequently diffuse staining (>50% of tubules or PTC) for both C5aR1 and C5aL2 than controls.

Article Snippet: After drying at room temperature, we fixed the samples in 10% buffered formalin for 10 min. After PBS washing, 3% goat serum in PBS was used to block nonspecific binding sites for 1 h. Primary antibodies against C5aR1 (#HM2094, Hycult Biotech) (mAb 1:10) and C5aL (#HP9036, Hycult Biotech) (pAb 1:25) were then incubated for one hour more.

Techniques: Staining, Transplantation Assay, Immunohistochemistry

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Promotion of C5aR-positive cancer cell invasion, tumor growth, and induction of MDSC recruitment by the Arthus reaction. Renca/C5aR cells (green) and Renca/mock cells (red) were labeled with different fluorescence probes, mixed, and injected subcutaneously into a wild-type mouse in combination with anti-ovalbumin mouse IgG or control IgG, followed by intravenous injection of ovalbumin (A, B) or with anti-BSA mouse IgG or control IgG, followed by intravenous injection of BSA (C-F). (A) Skin areas showing fluorescence from Renca/C5aR cells and Renca/mock cells (encircled in green and red lines, respectively), 24 h after ovalbumin injection. Scale bar: 200 µm. (B) Ratio of fluorescence distribution area of Renca/C5aR cells vs. that of Renca/mock cells was calculated. The values indicate the mean ± SD (n=4). Open circles: Anti-ovalbumin IgG; closed circles: Control IgG. Values were compared by analysis of variance (ANOVA) followed by Bonferroni multiple-comparison adjustment. *P=0.03, **P=0.067, ***P=0.008. (C) Images showing tumors at Renca cell-injected sites in the presence or absence of the Arthus reaction at day 14. Tumors are encircled in black lines. Scale bar: 1 cm. (D) Tumor growth at 7 and 14 days in sites subcutaneously injected with anti-BSA IgG (open column) or control IgG (closed column) after an intravenous BSA injection. Tumor size was measured and expressed as a ratio relative to the average tumor size in the absence of the Arthus reaction and vertical bars indicate the SD (n=6). *P=0.036, **P=0.021. (E) Fluorescence immunohistochemistry of Renca/C5aR cell-injected sites at day 14 using FITC-labeled anti-mouse CD11b rat IgG and Cy-5 ® -labeled anti-mouse Ly6g rat IgG. Upper and lower panels are from the sites where the Arthus reaction was induced or not, respectively. Scale bar: 50 µm. (F) MDSCs (CD11b + Ly6g + ) were counted in the cancer tissues where the Arthus reaction was induced (open column) or not (closed column) and the average MDSCs/high-power field (HPF) ± SD (n=6) were recorded. Values were compared by Wilcoxon rank-sum (Mann-Whitney) test for nonparameric distribution followed by Bonferroni multiple-comparison adjustment. *P=0.031, **P=0.031, n.s, not significant.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Labeling, Fluorescence, Injection, Immunohistochemistry, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Increase in lung nodules of C5aR-positive cancer cells stimulated with C5a and injected intravenously. (A) HuCCT1/C5aR cells incubated in the presence or absence of C5a or HuCCT1/mock cells incubated with C5a in the lung tissues of nude mice 6 weeks after cancer cell injection (hematoxylin-eosin stain). Nodules are encircled. Scale bar: 200 µm. The area in the square in the upper left panel is magnified in the lower left panel. Scale bar: 50 µm. (B) Lung tissues of nude mice six weeks after injection with cancer cells (Ki-67 immunohistochemical stain). Cancer nodules containing Ki-67-positive cells are encircled. HuCCT1/C5aR cells incubated in the presence (upper left) or absence (upper right) of C5a. HuCCT1/mock cells incubated with C5a (lower right). Scale bar: 200 µm. The area in the square in the upper left panel is magnified in the lower left panel. Scale bar: 200 µm. (C) Cancer nodules in five random microscopic fields (×100) in a section were counted, and the average colony number per field ± SD was recorded (n=6). *P<0.01, n.s, not significant.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Injection, Incubation, Staining, Immunohistochemical staining

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: C5aR expression in uterine cervical cells. Uterine cervical tissue samples were immunostained using an anti-human C5aR antibody. (A) Magnified image of the square marked in panel C. (B) Noncancerous epithelial cells. (C) Tissue with high C5aR expression. (D) Tissue with low C5aR expression. Scale bars: 50 µm in A and 100 µm in B-D.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Association between C5a receptor expression and invasion in uterus cervical cancer.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Promotion of C5aR-positive cancer cell invasion, tumor growth, and induction of MDSC recruitment by the Arthus reaction. Renca/C5aR cells (green) and Renca/mock cells (red) were labeled with different fluorescence probes, mixed, and injected subcutaneously into a wild-type mouse in combination with anti-ovalbumin mouse IgG or control IgG, followed by intravenous injection of ovalbumin (A, B) or with anti-BSA mouse IgG or control IgG, followed by intravenous injection of BSA (C-F). (A) Skin areas showing fluorescence from Renca/C5aR cells and Renca/mock cells (encircled in green and red lines, respectively), 24 h after ovalbumin injection. Scale bar: 200 µm. (B) Ratio of fluorescence distribution area of Renca/C5aR cells vs. that of Renca/mock cells was calculated. The values indicate the mean ± SD (n=4). Open circles: Anti-ovalbumin IgG; closed circles: Control IgG. Values were compared by analysis of variance (ANOVA) followed by Bonferroni multiple-comparison adjustment. *P=0.03, **P=0.067, ***P=0.008. (C) Images showing tumors at Renca cell-injected sites in the presence or absence of the Arthus reaction at day 14. Tumors are encircled in black lines. Scale bar: 1 cm. (D) Tumor growth at 7 and 14 days in sites subcutaneously injected with anti-BSA IgG (open column) or control IgG (closed column) after an intravenous BSA injection. Tumor size was measured and expressed as a ratio relative to the average tumor size in the absence of the Arthus reaction and vertical bars indicate the SD (n=6). *P=0.036, **P=0.021. (E) Fluorescence immunohistochemistry of Renca/C5aR cell-injected sites at day 14 using FITC-labeled anti-mouse CD11b rat IgG and Cy-5 ® -labeled anti-mouse Ly6g rat IgG. Upper and lower panels are from the sites where the Arthus reaction was induced or not, respectively. Scale bar: 50 µm. (F) MDSCs (CD11b + Ly6g + ) were counted in the cancer tissues where the Arthus reaction was induced (open column) or not (closed column) and the average MDSCs/high-power field (HPF) ± SD (n=6) were recorded. Values were compared by Wilcoxon rank-sum (Mann-Whitney) test for nonparameric distribution followed by Bonferroni multiple-comparison adjustment. *P=0.031, **P=0.031, n.s, not significant.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Labeling, Fluorescence, Injection, Immunohistochemistry, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Increase in lung nodules of C5aR-positive cancer cells stimulated with C5a and injected intravenously. (A) HuCCT1/C5aR cells incubated in the presence or absence of C5a or HuCCT1/mock cells incubated with C5a in the lung tissues of nude mice 6 weeks after cancer cell injection (hematoxylin-eosin stain). Nodules are encircled. Scale bar: 200 µm. The area in the square in the upper left panel is magnified in the lower left panel. Scale bar: 50 µm. (B) Lung tissues of nude mice six weeks after injection with cancer cells (Ki-67 immunohistochemical stain). Cancer nodules containing Ki-67-positive cells are encircled. HuCCT1/C5aR cells incubated in the presence (upper left) or absence (upper right) of C5a. HuCCT1/mock cells incubated with C5a (lower right). Scale bar: 200 µm. The area in the square in the upper left panel is magnified in the lower left panel. Scale bar: 200 µm. (C) Cancer nodules in five random microscopic fields (×100) in a section were counted, and the average colony number per field ± SD was recorded (n=6). *P<0.01, n.s, not significant.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Injection, Incubation, Staining, Immunohistochemical staining

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: C5aR expression in uterine cervical cells. Uterine cervical tissue samples were immunostained using an anti-human C5aR antibody. (A) Magnified image of the square marked in panel C. (B) Noncancerous epithelial cells. (C) Tissue with high C5aR expression. (D) Tissue with low C5aR expression. Scale bars: 50 µm in A and 100 µm in B-D.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Enhancement of cancer invasion and growth via the C5a-C5a receptor system: Implications for cancer promotion by autoimmune diseases and association with cervical cancer invasion

doi: 10.3892/ol.2018.9715

Figure Lengend Snippet: Association between C5a receptor expression and invasion in uterus cervical cancer.

Article Snippet: Recombinant human C5a and anti-human C5aR mouse monoclonal IgG were purchased from EMD Millipore (Billerica, MA, USA) and Hycult Biotech (Uden, The Netherlands), respectively.

Techniques: Expressing