rabbit primary antibodies  (Novus Biologicals)

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: CFA induces mechanical hyperalgesia and alters spinal cord protein expression in rats. (a) Paw withdrawal thresholds (PWTs) of rats in each group on different days ( n = 6). (b) Western blot analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord of vehicle-injected and CFA-injected rats on days −1, 1, 3, 5, 7, 10, and 14 after CFA injection. (c) Quantitative analysis of the relative expressions of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cords of vehicle-injected and CFA-injected rats on days 1, 3, 7, and 14 after CFA injection ( n = 6). * p < 0.05, ** p < 0.01 versus vehicle-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Expressing, Western Blot, Injection

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Endogenous expression and cellular localization of CCL2 and CCR2 in the spinal cord of CFA-injected rats. (a) and (b) Double-label immunofluorescence assays were used to detect the colocalization of CCL2 and CCR2 with NeuN, Iba1, and GFAP in CFA-injected rats. White arrows indicate the colocalization of CCL2 and CCR2 with GFAP and NeuN. Scale bar = 100 μm.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Expressing, Injection, Immunofluorescence

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of the AP-1 inhibitor T-5224 attenuates CFA-induced inflammatory pain and suppresses the expression of c-Jun, p-c-Jun, CCL2, and CCR2. (a) Effect of intrathecal injection of T-5224 on PWT ( n = 6). (b) Western blot analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in spinal cords following intrathecal injection. (c) Quantitative analysis of the relative expressions of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). T-5224 (500 μg/20 μL, i.t.) was administered once daily on days 2, 3, and 4 after CFA injection. PWTs were measured 4 h after each injection. L4-L6 spinal tissues were collected 4 h after the last injection. * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Western Blot

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of the CCR2 antagonist RS 504393 attenuates CFA-induced inflammatory pain and suppresses the expression of CCL2 and CCR2. (a) Effect of intrathecal injection of RS 504393 on PWT ( n = 6). (b) Western blot analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in spinal cords following intrathecal injection. (c) Quantitative analysis of the relative expressions of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). RS 504393 (20 μg/20 μL, i.t.) was administered once daily on days 2, 3, and 4 after CFA injection. PWTs were measured 4 h after each injection. L4-L6 spinal tissues were collected 4 h after the last injection. * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Western Blot

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of the AP-1 inhibitor T-5224 suppresses the expression of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord. (a) Immunofluorescence analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord following intrathecal injection. (b) Mean fluorescence intensity of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Immunofluorescence, Fluorescence

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of the CCR2 antagonist RS 504393 suppresses the expression of CCL2 and CCR2 in the spinal cord. (a) Immunofluorescence analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord following intrathecal injection. (b) Mean fluorescence intensity of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Immunofluorescence, Fluorescence

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of recombinant rat CCL2 protein enhances CFA-induced inflammatory pain and increases the expression of CCL2 and CCR2, whereas T-5224 treatment reverses these effects. (a) Repeated intrathecal injections of recombinant rat CCL2 protein significantly increased mechanical pain in rats compared to the CFA-injected group, while T-5224 treatment reversed this effect ( n = 6). (b) Western blot analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in spinal cords following intrathecal injection. (c) Quantitative analysis of the relative expressions of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). Recombinant rat CCL2 protein (1 µg/10 µL, i.t.) or T-5224 (500 µg/20 µL, i.t.) was administered once daily on days 2, 3, and 4 after CFA injection. PWTs were measured 4 h after each injection. L4-L6 spinal tissues were collected 4 h after the last injection. * p < 0.05, ** p < 0.01 versus CFA + CCL2 group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Recombinant, Expressing, Western Blot

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Mean fluorescence intensity of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). * p < 0.05, ** p < 0.01 versus CFA + CCL2; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Fluorescence, Injection

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of curcumin attenuates CFA-induced inflammatory pain and suppresses the expression of c-Jun, p-c-Jun, CCL2, and CCR2. (a) Effect of intrathecal injection of curcumin on paw withdrawal thresholds (PWTs; n = 6). (b) Western blot analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in spinal cords following intrathecal injection. (c) Quantitative analysis of the relative expressions of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). Curcumin (100 µg/20 µL, 500 µg/20 µL, and 1 mg/20 µL, i.t.) was administered once daily on days 2, 3, and 4 after CFA injection. PWTs were measured 4 h after each injection. L4-L6 spinal tissues were collected 4 h after the last injection. * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Western Blot

Journal: Molecular Pain

Article Title: Curcumin relieves CFA-induced inflammatory pain by inhibiting the AP-1/c-Jun-CCL2-CCR2 pathway in the spinal dorsal horn

doi: 10.1177/17448069251323668

Figure Lengend Snippet: Intrathecal injection of curcumin suppresses the expression of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord. (a) Immunofluorescence analysis of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord following intrathecal injection. (b) Mean fluorescence intensity of c-Jun, p-c-Jun, CCL2, and CCR2 in the spinal cord ( n = 6). * p < 0.05, ** p < 0.01 versus vehicle-injected group; # p < 0.05, ## p < 0.01 versus CFA-injected group.

Article Snippet: Primary antibodies were then incubated overnight at 4°C: rabbit anti-c-Jun antibody (9165S, Cell Signaling, 1:1000), rabbit anti-p-c-Jun antibody (91952S, Cell Signaling, 1:1000), rabbit anti-CCR2 antibody (16153-1-AP, Proteintech, 1:1000), rabbit anti-CCL2 antibody (26161-1-AP, Proteintech, 1:500), and mouse anti-tubulin antibody (1:3000).

Techniques: Injection, Expressing, Immunofluorescence, Fluorescence

Journal: bioRxiv

Article Title: Interleukin-1β Drives Disease Progression in Arrhythmogenic Cardiomyopathy

doi: 10.1101/2024.12.11.628020

Figure Lengend Snippet: ( A ) UMAP of fibroblast populations. ( B ) Composition plots of fibroblast populations between Donor controls and ACM. ( C ) Major fibroblast gene markers overlaid on the fibroblast UMAP. ( D ) Heatmap displaying expression of fibroblast markers across spatial niches. ( E ) Fibroblast differential gene expression signature associated with either Donor controls or ACM overlaid onto the fibroblast UMAP space. ( F ) ACM fibroblast gene expression signature overlaid onto ACM tissue space. ( G ) UMAP of myeloid populations. ( H ) Composition plots of myeloid populations between Donor controls and ACM. ( I ) Major gene markers for myeloid populations overlaid on the myeloid UMAP. ( J ) Pathway analysis displaying pathways upregulated in ACM relative to Donor controls based on differentially expressed myeloid genes. ( K ) Colocalization of the inflammatory macrophage population (Mac4) and the POSTN+ fibroblast population (Fib3) in areas of tissue damage and fibrosis and heatmap displaying expression of major inflammatory genes across spatial niches. ( L ) Immunofluorescence staining displaying colocalization of CCR2+ CD68+ macrophages and FAP+ fibroblasts in ACM tissue samples. Two independent samples were used. Broader images are captured at 20x.

Article Snippet: Slides were then washed in 1X PBS (3x, 5mins/wash), incubated with 5% donkey serum (Stratech Scientific, Cat. No. 017-000-121), 1% BSA (Merck Life Science, Cat. No. A2153) and 0.15% Triton-X (ThermoFisher, Cat. No. A16046.0F) in 1X PBS for 1hr and then probed with rat anti-CD68 antibody (ThermoFisher, Cat. No. 14-0681-82, at 1:200) and rabbit anti-CCR2 antibody (ThermoFisher, Cat. No. BS-23026R, at 1:400) overnight at 4°.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Interleukin-1β Drives Disease Progression in Arrhythmogenic Cardiomyopathy

doi: 10.1101/2024.12.11.628020

Figure Lengend Snippet: IL1β inhibition prevents NFĸB nuclear localization in cardiac myocytes and infiltrating myocardial CCR2/CD68+ macrophages in Dsg2 mut/mut mice. ( A ) Representative immunostained hearts probed for RelA, CCR2/CD68, JUP, and Cx43. n≥5 hearts/cohort/stain; black arrows, cardiomyocyte RelA positive nuclei; red arrowheads, CCR2/CD68+ macrophages; red arrows: CCR2/CD68+ cardiac myocytes. ( B ) Number of cells per mm 2 positive for nuclear RelA localization. ( C ) Number of cells per mm 2 positive for CCR2. ( D ) Pearson’s correlation analysis for cells that showed dual labeling for RelA and CCR2 (P- and r-values inset). For B and C, *P<0.05 for any cohort vs WT+Isotype; and † P<0.05 for any cohort vs Dsg2 mut/mut +Isotype

Article Snippet: Slides were then washed in 1X PBS (3x, 5mins/wash), incubated with 5% donkey serum (Stratech Scientific, Cat. No. 017-000-121), 1% BSA (Merck Life Science, Cat. No. A2153) and 0.15% Triton-X (ThermoFisher, Cat. No. A16046.0F) in 1X PBS for 1hr and then probed with rat anti-CD68 antibody (ThermoFisher, Cat. No. 14-0681-82, at 1:200) and rabbit anti-CCR2 antibody (ThermoFisher, Cat. No. BS-23026R, at 1:400) overnight at 4°.

Techniques: Inhibition, Staining, Labeling