anti inos antibody  (R&D Systems)

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