antibodies against human inos  (Servicebio Inc)

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: In vitro assay of inflammation cell modulation under stimulation of SP-loaded Gel/HA and IL-10-loaded Ker/Cu. a Schematic of neutrophil migration test using a transwell system after treatment with the leaching solution of SP@Gel/HA. Gel/HA and blank culture medium were set for comparison. b Wright-Giemsa staining of HL-60 cells before and after differentiation. c Photograph of dHL-60 cells migrating to the lower chamber. d Quantitative analysis of neutrophil migration after treatments with SP@Gel/HA and Gel/HA. Untreated group serves as a control. e Schematic of macrophage polarization after treatment with LPS, IL-10, or IL-10/LPS. f Representative fluorescence images of macrophages after different treatment. Red: iNOS (M1 marker); Green: CD163 (M2c marker); Blue: DAPI (nuclear staining). g Schematic of macrophage efferocytosis test toward apoptotic dHL-60 cells under different treatments. h Flow cytometry plots of dHL-60 cells before and after apoptosis induction. i Representative fluorescent images of macrophage efferocytosis toward apoptotic dHL-60 cells under different treatments. Macrophages and apoptotic cells were stained green and red, respectively. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. ns, not significant; ∗∗∗∗p < 0.0001.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: In Vitro, Migration, Comparison, Staining, Control, Fluorescence, Marker, Flow Cytometry, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation

Journal: Scientific Reports

Article Title: Thrombin induces degradation of murine intervertebral discs via angiogenesis and M1-like polarization of macrophages

doi: 10.1038/s41598-025-18684-6

Figure Lengend Snippet: Thrombin promotes polarization into M1 macrophages. ( a ) Mouse bone marrow was stimulated with thrombin, and quantitative PCR was performed using specific primers for CD86 and CD163 . Biological and technical replicates were performed three times. ( b ) Mouse bone marrow was stimulated with thrombin, and western blotting was performed using antibodies for the M1 markers iNOS and CD86, and the M2 markers CD163 and Arginase-1. β-actin was used as an internal control. n = 3. Values represent the mean ± SD. * p < 0.05, ** p < 0.01, relative to the corresponding control.

Article Snippet: Equal amounts of protein from each sample were analyzed, as previously described , using immunoblotting with primary antibodies against iNOS (MAB9502, 1:500, R&D Systems), CD86 (NBP2-25208, 1:250, Novus Biologicals), Arginase-1 (#93668, 1:1000, Cell Signaling Technology), CD163 (ab182422, 1:1000, Abcam), and β-actin (A2228, 1:1000, Sigma-Aldrich). β-actin was used for normalization of relative protein levels.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Scientific Reports

Article Title: Thrombin induces degradation of murine intervertebral discs via angiogenesis and M1-like polarization of macrophages

doi: 10.1038/s41598-025-18684-6

Figure Lengend Snippet: M1 macrophages were induced in the in vivo mouse intervertebral disc (mIVD) degeneration model (puncture model). ( a ) Immunohistological analysis of Iba-1, iNOS, and arginase (brown staining) expression in nucleus pulposus (NP), annulus fibrosis (AF), and cartilage endplate (CEP) [right] at high magnification and in whole mIVDs (left) at low magnification. Images using an IgG isotype control antibody are included to demonstrate the specificity of the IHC staining. ( b ) Protein was extracted from punctured mIVDs in the puncture model, and western blotting was performed using antibodies against CD86, CD163, and β-actin. ( c ) Images of the panel in ( b ), captured using a ChemiDoc Touch system and quantified using ImageJ. Biological and technical replicates were performed three times. Values represent the mean ± SD. * p < 0.05, ** p < 0.01, relative to the corresponding control.

Article Snippet: Equal amounts of protein from each sample were analyzed, as previously described , using immunoblotting with primary antibodies against iNOS (MAB9502, 1:500, R&D Systems), CD86 (NBP2-25208, 1:250, Novus Biologicals), Arginase-1 (#93668, 1:1000, Cell Signaling Technology), CD163 (ab182422, 1:1000, Abcam), and β-actin (A2228, 1:1000, Sigma-Aldrich). β-actin was used for normalization of relative protein levels.

Techniques: In Vivo, Staining, Expressing, Control, Immunohistochemistry, Western Blot

Journal: Scientific Reports

Article Title: Thrombin induces degradation of murine intervertebral discs via angiogenesis and M1-like polarization of macrophages

doi: 10.1038/s41598-025-18684-6

Figure Lengend Snippet: PAR1 antagonist abrogates macrophage M1 polarization and angiogenesis. Intervertebral disc tissue was obtained from the puncture model 3 weeks after puncture, with or without PAR1-antagonist treatment. Immunostaining was performed using antibodies against CD86, iNOS, CD163, arginase, and thrombin. Images using an IgG isotype control antibody are included to demonstrate the specificity of the IHC staining. Three mice were used in each group.

Article Snippet: Equal amounts of protein from each sample were analyzed, as previously described , using immunoblotting with primary antibodies against iNOS (MAB9502, 1:500, R&D Systems), CD86 (NBP2-25208, 1:250, Novus Biologicals), Arginase-1 (#93668, 1:1000, Cell Signaling Technology), CD163 (ab182422, 1:1000, Abcam), and β-actin (A2228, 1:1000, Sigma-Aldrich). β-actin was used for normalization of relative protein levels.

Techniques: Immunostaining, Control, Immunohistochemistry

Journal: Scientific Reports

Article Title: Fisetin inhibits lipopolysaccharide-induced inflammatory response by activating β-catenin, leading to a decrease in endotoxic shock

doi: 10.1038/s41598-021-87257-0

Figure Lengend Snippet: Fisetin inhibits LPS-induced inflammatory response in zebrafish larvae. Zebrafish larvae at 1 day post fertilization (dpf) were cultured in 0.003% PTU containing E3 embryo media. Briefly, 2 nL of 0.5 mg/mL LPS was microinjected into the yolk at 3 dpf. Zebrafish larvae were immediately immersed in E3 embryo media containing different concentrations of fisetin. ( A ) In LPS-microinjected conditions, 10 zebrafish were euthanized at the indicated time points and subjected to RT-PCR for evaluating the expression of iNOS , COX-2a, IL-6 , and TNF-α . ( B ) At 18 h post injection (hpi), 20 zebrafish larvae from each treatment were euthanized and the expression of iNOS , COX-2a, IL-6 , and TNF-α was measured by RT-PCR. ( C ) Neutral red staining of macrophages and ( D ) sudan black staining of the neutrophils were performed at 24 hpi. Each value indicates the mean ± standard error median (SEM) and is representative of the results obtained from three independent experiments. Significant differences among the groups were determined using the Student’s t -test ( ### p < 0.001, ## p < 0.01, and # p < 0.05 vs. untreated zebrafish larvae) and one-way ANOVA with Bonferroni correction (*** p < 0.001, ** p < 0.01, and * p < 0.05 vs. LPS-treated zebrafish larvae).

Article Snippet: Mouse anti-human antibodies against iNOS (sc-7271), COX-2 (sc-19999), p50 (sc-8414), p65 (sc-8008), GSK-3β (sc-81462), phospho (p)-GSK-3β at Ser9 (sc-37800), β-catenin (sc-59737), β-actin (sc-69879), nucleolin (sc-13057), and peroxidase-labeled anti-mouse immunoglobulins were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection, Staining

Journal: Scientific Reports

Article Title: Fisetin inhibits lipopolysaccharide-induced inflammatory response by activating β-catenin, leading to a decrease in endotoxic shock

doi: 10.1038/s41598-021-87257-0

Figure Lengend Snippet: Fisetin inhibits LPS-induced inflammation and endotoxic shock by stimulating the β-catenin signaling pathway in zebrafish larvae. Zebrafish larvae at 3 dpf were pretreated with 10 µM FH535 for 2 h prior to LPS (2 nL of 0.5 mg/mL) microinjection. Then, the larvae were grown in E3 media containing 400 μM fisetin. ( A ) Neutral red-stained macrophages are shown at 18 hpi. ( B ) Sudan black-stained neutrophils were observed at 18 hpi. ( C ) At 18 hpi, 10 zebrafish from each condition were collected and the expression of iNOS and COX-2a was detected by RT-PCR. GAPDH was used as an internal control. Each value indicates the mean ± standard error median (SEM) and is representative of the results obtained from three independent experiments. Significant differences among the groups were determined using one-way ANOVA with Bonferroni correction (*** p < 0.001, ** p < 0.005, and * p < 0.01).

Article Snippet: Mouse anti-human antibodies against iNOS (sc-7271), COX-2 (sc-19999), p50 (sc-8414), p65 (sc-8008), GSK-3β (sc-81462), phospho (p)-GSK-3β at Ser9 (sc-37800), β-catenin (sc-59737), β-actin (sc-69879), nucleolin (sc-13057), and peroxidase-labeled anti-mouse immunoglobulins were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Microinjection, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: Fisetin inhibits lipopolysaccharide-induced inflammatory response by activating β-catenin, leading to a decrease in endotoxic shock

doi: 10.1038/s41598-021-87257-0

Figure Lengend Snippet: Fisetin inhibits GSK-3β-mediated NF-κB activation in the presence of β-catenin, leading to the inhibition of inflammation-induced septic shock. Once macrophages are exposed to high concentrations of the bacterial endotoxin, LPS, they initiate an inflammatory response and endotoxic shock by upregulating the expression of NF-κB-induced inflammatory genes, such as iNOS, COX-2, IL-12, IL-6, and TNF-α . Fisetin binds to the noncompetitive ATP-binding sites of GSK-3β and phosphorylates GSK-3β at Ser9, resulting in the inactivation GSK-3β and release of β-catenin from the destruction complex. The released β-catenin inhibits the transcriptional activity of NF-κB, thereby alleviating LPS-induced inflammation and endotoxic shock.

Article Snippet: Mouse anti-human antibodies against iNOS (sc-7271), COX-2 (sc-19999), p50 (sc-8414), p65 (sc-8008), GSK-3β (sc-81462), phospho (p)-GSK-3β at Ser9 (sc-37800), β-catenin (sc-59737), β-actin (sc-69879), nucleolin (sc-13057), and peroxidase-labeled anti-mouse immunoglobulins were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activation Assay, Inhibition, Expressing, Binding Assay, Activity Assay