mouse antibodies against ccl2  (Bio X Cell)

Journal: Cancer Research Communications

Article Title: Defining the Ovarian Cancer Precancerous Landscape through Modeling Fallopian Tube Epithelium Reprogramming Driven by Extracellular Vesicles

doi: 10.1158/2767-9764.CRC-25-0064

Figure Lengend Snippet: Validation of 1-day and 14-day treatment transcriptomics data using IHC. A, Left, CCL2 transcript expression in OVCAR3-, FT240-, and PBS-treated secretory cells after 1 day. Right, IHC staining for upregulated CCL2 in FTE from three individual patients. B, Left, FLNA transcript expression in OVCAR3-, FT240-, and PBS-treated secretory cells after 1 day. Right, IHC staining for downregulated FLNA in FTE from three individual patients. C, Left, VCAM1 transcript expression in OVCAR3-, FT240-, and PBS-treated secretory cells after 1 day. Right, IHC staining for upregulated VCAM1 in FTE from three individual patients. D, Left, TPI1 transcript expression in OVCAR3-, FT240-, PBS-treated secretory cells after 1 day. Right, IHC staining for downregulated protein TPI1 in FTE from three individual patients. E, Left, TXNIP transcript expression in OVCAR3-, FT240-, and PBS-treated secretory cells after 14 days. Right, IHC staining for downregulated TXNIP in FTE from three individual patients. F, Left, VCAM1 transcript expression in OVCAR3-, FT240-, and PBS-treated ciliated cells after 14 days. Right, IHC staining for upregulated VCAM1 in FTE from three individual patients. In all boxplots, each point represents normalized transcript abundance measured in a single segment. Scale bar, 20 μm; arrows indicate epithelium.

Article Snippet: Primary antibodies against CCL2 (Novus, cat. #NBP1-07035SS, RRID: AB_1625611), VCAM1 (Thermo Fisher Scientific, cat. #MA5-31965, RRID: AB_2809259), FLNA (Proteintech, cat. #67133-1-Ig, RRID: AB_2882432), TPI1 (Proteintech, cat. #10713-1-AP, RRID: AB_2207716), and TXNIP (Thermo Fisher Scientific, cat. #40-3700, RRID: AB_2533462) were incubated overnight at 4°C, followed by detection using a horseradish peroxidase–linked secondary antibody and DAB substrate (the full procedure is provided in the Supplementary Methods).

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Antifibrotic effects of specific targeting of the 5‐hydroxytryptamine 2B receptor (5‐HT 2B R) in murine models and ex vivo models of scleroderma skin

doi: 10.1002/art.43151

Figure Lengend Snippet: Inhibition of 5‐HT 2B R by AM1476 modulates the expression of MAOB, SCL6A4, CCL2, CCL5, PAI‐1, and P‐STAT3 in murine cGvHD and PCS slices of SSc skin. (A and B) Murine cGvHD model. (A) Representative images of immunofluorescence staining and (B) quantification of Maob, Slc6a4, Ccl2, CCl5, Pai‐1, and P‐Stat3 in the skin of syngeneically and allogeneically transplanted mice with or without AM1476 treatment. All data are presented as mean ± SEM, with individual values displayed as column plus dots. Differences between the groups were tested for their statistical significance by one‐way analysis of variance with Dunnett's multiple comparison. Adjusted P values less than 0.05 were considered significant. Adjusted P values are expressed as follows: *0.05 > P > 0.01; **0.01 > P > 0.001; ***0.001 > P > 0.0001; **** P < 0.0001 as compared to the control allogenic group. (C and D) PCS slices of SSc skin. (C) Representative immunofluorescence staining images and (D) quantification of MAOB, SLC6A4, CCL2, CCL18, PAI‐1, and P‐STAT3 in SSc‐PCS with or without AM1476. All data points are presented as individual values displayed as dots. Differences between the groups were tested for their statistical significance by two‐way analysis of variance with Sidak's multiple comparison. Adjusted P values less than 0.05 were considered significant. Adjusted P values are expressed as follows: *0.05 > P > 0.01; **0.01 > P > 0.001; ***0.001 > P > 0.0001; **** P < 0.0001 as compared to the SSc‐PCS control group. 5‐HT 2B R, 5‐hydroxytryptamine 2B receptor; cGvHD, chronic graft‐versus‐host disease; PCS, precision cut skin; SSc, systemic sclerosis. Color figure can be viewed in the online issue, which is available at http://onlinelibrary.wiley.com/doi/10.1002/art.43151/abstract .

Article Snippet: The staining of skin sections was performed by using the antibodies against CCL2 (LS‐C169178‐100, LSBio, 1:100 dilution), CCL5 (MAB478‐100, Biotechne, 1:100 dilution), CCL18 (22303‐1‐AP, Proteintech, 1:100 dilution), PAI‐1 (66261‐1‐Ig, Proteintech, 1:100 dilution), P‐STAT3 (MA5‐15193, Thermo Scientific, 1:100 dilution), MAOB (12602‐1‐AP, Proteintech, 1:100 dilution), or SLC6A4 (LS‐C154958‐100, LSBio, 1:100 dilution).

Techniques: Inhibition, Expressing, Immunofluorescence, Staining, Comparison, Control

Journal: Brain research bulletin

Article Title: Neuroprotective effects of hirudin against cerebral ischemia-reperfusion injury via inhibition of CCL2-mediated ferroptosis and inflammatory pathways.

doi: 10.1016/j.brainresbull.2025.111293

Figure Lengend Snippet: Fig. 4 CCL2 overexpression partially reverses the protective effect of hirudin on

Article Snippet: Brain tissues were fixed within 4% paraformaldehyde and sliced, followed by blocking and an overnight incubation at 4°C with primary antibody against CCL2 (1:50, 66272-1-Ig, Proteintech).

Techniques: Over Expression

Journal: Brain research bulletin

Article Title: Neuroprotective effects of hirudin against cerebral ischemia-reperfusion injury via inhibition of CCL2-mediated ferroptosis and inflammatory pathways.

doi: 10.1016/j.brainresbull.2025.111293

Figure Lengend Snippet: Fig. 5 CCL2 overexpression partially reverses the protective effect of Hirudin on

Article Snippet: Brain tissues were fixed within 4% paraformaldehyde and sliced, followed by blocking and an overnight incubation at 4°C with primary antibody against CCL2 (1:50, 66272-1-Ig, Proteintech).

Techniques: Over Expression

Journal: Brain research bulletin

Article Title: Neuroprotective effects of hirudin against cerebral ischemia-reperfusion injury via inhibition of CCL2-mediated ferroptosis and inflammatory pathways.

doi: 10.1016/j.brainresbull.2025.111293

Figure Lengend Snippet: Fig. 6 Hirudin inhibits CCL2-mediated neuronal ferroptosis in OGD/R neurons

Article Snippet: Brain tissues were fixed within 4% paraformaldehyde and sliced, followed by blocking and an overnight incubation at 4°C with primary antibody against CCL2 (1:50, 66272-1-Ig, Proteintech).

Techniques:

Journal: Brain research bulletin

Article Title: Neuroprotective effects of hirudin against cerebral ischemia-reperfusion injury via inhibition of CCL2-mediated ferroptosis and inflammatory pathways.

doi: 10.1016/j.brainresbull.2025.111293

Figure Lengend Snippet: Fig. 8 Hirudin inhibits CCL2-mediated activation of the TLR4/NF-κB signaling

Article Snippet: Brain tissues were fixed within 4% paraformaldehyde and sliced, followed by blocking and an overnight incubation at 4°C with primary antibody against CCL2 (1:50, 66272-1-Ig, Proteintech).

Techniques: Activation Assay

Journal: Stem Cells Translational Medicine

Article Title: Mitigating murine acute and chronic Coxsackievirus B3‐induced myocarditis with human right atrial appendage-derived stromal cells

doi: 10.1093/stcltm/szae103

Figure Lengend Snippet: RAA-CardAPs modulate cardiac chemokine expression and splenic pro- and anti-inflammatory monocytes in acute CVB3-induced myocarditis mice. (A-E) The impact of RAA-CardAPs and EMB-CardAPs on LV protein expression of the chemokines CCL2 and CCL7 attracting pro-inflammatory monocytes, and CX3CL1, attracting anti-inflammatory monocytes in PBS-injected and CVB3-infected mice was analyzed via LV immunohistological stainings of (A) CCL2, (B) CCL7, (C) CX3CL1, depicted as positive area (%)/HA (mm 2 ). (D-F) To determine the impact of RAA-CardAPs and EMB-CardAPs on splenic pro-inflammatory and anti-inflammatory monocytes, the percentage of (D) pro-inflammatory Ly6C high CCR2 high CX3CR1 low and (E) Ly6C mid CCR2 high CX3CR1 low of gated CD115 + CD11b + and the percentage of (F) anti-inflammatory Ly6C low CCR2 low CX3CR1 high of gated CD115 + CD11b + monocytes in the spleen was analyzed. Data are represented as scatter plots with bars, showing individual data points and the corresponding mean ± SEM. Statistical differences were assessed using 1-way ANOVA or Kruskal-Wallis test (* P < .05, ** P < .01, *** P < 0.001, and P < .0001, n = 5/group for all analysis except for (D-F) n = 3-4/group).

Article Snippet: Frozen LV tissue samples were cut in 5 μm thick sections and used for immunohistological staining with the following antibodies directed against CCL2 (Abnova, Taipei, Taiwan, PAB16617, 1:25, overnight), CCL7 (Cloud clone corp., Katy, TX, USA, PAA089Mu01, 1:25, overnight), CX3CL1 (Abcam, Cambridge, UK, AB25088, 1:75), collagen I (Chemicon, Darmstadt, Germany, AB765P, 1:300) and CD68 (Abcam, Cambridge, UK, AB53444, 1:600).

Techniques: Expressing, Injection, Infection

Journal: Stem Cells Translational Medicine

Article Title: Mitigating murine acute and chronic Coxsackievirus B3‐induced myocarditis with human right atrial appendage-derived stromal cells

doi: 10.1093/stcltm/szae103

Figure Lengend Snippet: RAA-CardAPs decrease splenic pro-inflammatory monocytes in chronic CVB3-induced myocarditis mice. (A-C) LV mRNA expression of the chemokines CCL2, CCL7, and CX3CL1 was analyzed via real-time PCR with (A) CCL2, (B) CCL7, and (C) CX3CL1 mRNA expression depicted as n -fold with control mice set as 1. (D-F) The impact of RAA-CardAPs injected at d10 post CVB3 infection on splenic pro-inflammatory and anti-inflammatory monocytes was evaluated. Therefore, the percentage of (D) pro-inflammatory Ly6C high CCR2 high CX3CR1 low and (E) Ly6C mid CCR2 high CX3CR1 low of gated CD115 + CD11b + and the percentage of (F) anti-inflammatory Ly6C low CCR2 low CX3CR1 high of gated CD115 + CD11b + monocytes in the spleen was analyzed. Data are represented as scatter plots with bars, showing individual data points and the corresponding mean ± SEM. Statistical differences were assessed using 1-way ANOVA or Kruskal-Wallis test (* P < .05, ** P < .01, *** P < .001, and **** P < .0001, (A-C) n = 9/Control and Control + RAA-CardAPs, n = 14-17/CVB3, and CVB3 + RAA-CardAPs, (D-F) n = 5-9/group).

Article Snippet: Frozen LV tissue samples were cut in 5 μm thick sections and used for immunohistological staining with the following antibodies directed against CCL2 (Abnova, Taipei, Taiwan, PAB16617, 1:25, overnight), CCL7 (Cloud clone corp., Katy, TX, USA, PAA089Mu01, 1:25, overnight), CX3CL1 (Abcam, Cambridge, UK, AB25088, 1:75), collagen I (Chemicon, Darmstadt, Germany, AB765P, 1:300) and CD68 (Abcam, Cambridge, UK, AB53444, 1:600).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Injection, Infection

Journal: Nature Communications

Article Title: Migration arrest and transendothelial trafficking of human pathogenic-like Th17 cells are mediated by differentially positioned chemokines

doi: 10.1038/s41467-025-57002-6

Figure Lengend Snippet: a Culture medium collected from the upper and lower wells of transwells with HUVEC monolayers either not stimulated (NS) or stimulated with TNF-α + IFN-γ were assayed for CCL2, CCL7, and CCL8 by ELISA ( n = 3 separate experiments). Each symbol shows data from one experiment, with assays done in duplicate, and bars indicate means ± SEM. b Confocal microscopy images at × 40 magnification of permeabilized and non-permeabilized TNF-α + IFN-γ-stimulated HUVECs immunostained for CCL2, CCL7, CCL8, CCL5, CCL20, and CXCL9 (green) and stained using phalloidin for polymerized actin (magenta) and DAPI for nuclei (blue). The scale bars indicate 10 µm. Images are representative of three experiments. c Biotinylated CCL5 (0.1 μM) and CCL2 (1 μM) were incubated with CHO-K1 cells and the heparan-sulfate deficient cell line, D-677. Chemokine binding was detected using streptavidin-phycoerythrin (PE) and flow cytometry. Data shown are representative of three separate experiments.

Article Snippet: Samples were incubated at room temperature for 2 h with primary mouse antibodies against human chemokines CCL2 (5 μg, Cat# MAB679) CCL5 (5 μg, Cat# MAB278), CCL7 (5 μg, Cat# MAB282), CCL8 (5 μg, Cat# MAB281), CCL20 (5 μg, Cat# AF360), CXCL9 (5 μg, Cat# MAB392) from R&D Systems or with mouse IgG 1 isotype control (5 μg, Cat# MAB002) or mouse IgG 2B isotype control (Cat# MAB004) from R&D Systems, or with anti-human GOLPH2 (2 μg, Cat# PA5-30622) or rabbit IgG isotype control from Invitrogen, or with anti-human vWF (2 μg, Cat# ab6994) from Abcam.

Techniques: Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining, Incubation, Binding Assay, Flow Cytometry

Journal: Nature Communications

Article Title: Migration arrest and transendothelial trafficking of human pathogenic-like Th17 cells are mediated by differentially positioned chemokines

doi: 10.1038/s41467-025-57002-6

Figure Lengend Snippet: For ( a , c ), human CD4 + T cell subgroups were isolated by FACS from the blood of healthy donors as in Supplementary Fig. and Fig. . a CCR6 +(high) CCR2 + cells were either left untreated (marked with a minus sign) or treated with CCL2 (100 ng/ml) just before and for four min after adding the cells to the flow chambers, and CCL2-treated cells were used without (marked with a minus sign) or with treatment with the CCR2 antagonist, BMS22. Data points in the left panel show numbers of cells rolling, arrested, and transmigrated, while the middle panel shows arrested cells as percentages of cells rolling and the right panel shows transmigrated cells as percentages of cells arresting. Each symbol shows data from one donor, with n = 4 individual donors in four separate experiments using CCL2. Two of these experiments included the additional treatment with BMS22. b Above is the CCL2-CXCL9 chimera sequence with signal peptide sequence shown in green, CCL2-sequence in black, and C-terminal GAG-binding sequence of CXCL9 in red. Below are confocal microscopy images of control or CCL2-CXCL9-chimera transduced, TNF-α-stimulated HUVECs immunostained for CCL2 (red) and stained with DAPI for nuclei (blue). Images are representative of three experiments. c Numbers of CCR6 +(high) CCR2 + cells, either untreated (marked with a minus sign) or treated with the CCR2 antagonist, BMS22, rolling, arrested, and transmigrated on TNF- α -activated HUVECs transduced with either control virus (marked with a minus sign) or virus encoding the CCL2-CXCL9 chimera. Middle panel shows arrested cells as percentages of cells rolling and in the right panel transmigrated cells as percentages of cells arresting. Each symbol shows data from one donor, with cells from four donors and separate experiments in each treatment group except for the BMS22-treated cells with control virus-transduced HUVECs, where cells from three donors were used. Bars indicate means ± SEM. p values were calculated using two-tailed paired Student’s t tests, and no corrections were made for multiple comparisons. Source data are provided in the file, and representative videos used to quantify rolling, arrest, and transendothelial migration are provided in Supplementary Movies – .

Article Snippet: Samples were incubated at room temperature for 2 h with primary mouse antibodies against human chemokines CCL2 (5 μg, Cat# MAB679) CCL5 (5 μg, Cat# MAB278), CCL7 (5 μg, Cat# MAB282), CCL8 (5 μg, Cat# MAB281), CCL20 (5 μg, Cat# AF360), CXCL9 (5 μg, Cat# MAB392) from R&D Systems or with mouse IgG 1 isotype control (5 μg, Cat# MAB002) or mouse IgG 2B isotype control (Cat# MAB004) from R&D Systems, or with anti-human GOLPH2 (2 μg, Cat# PA5-30622) or rabbit IgG isotype control from Invitrogen, or with anti-human vWF (2 μg, Cat# ab6994) from Abcam.

Techniques: Isolation, Sequencing, Binding Assay, Confocal Microscopy, Control, Staining, Transduction, Virus, Two Tailed Test, Migration

Journal: Nature Communications

Article Title: Migration arrest and transendothelial trafficking of human pathogenic-like Th17 cells are mediated by differentially positioned chemokines

doi: 10.1038/s41467-025-57002-6

Figure Lengend Snippet: Cytokine-activated endothelial cells upregulate expression of selectins and integrin ligands and secrete chemokines. Selectin-selectin ligand interactions help capture cells and allow them to roll along the endothelium. For pathogenic-like type 17 cells, endothelial cell-bound chemokines can then stimulate CCR6, CCR5, and CXCR3 to activate integrins for binding to intercellular adhesion molecules and mediate firm arrest. Secreted chemokines that fail to adhere to endothelial cells, such as CCL2 and other CCR2 ligands, are removed by blood flow to create a transendothelial gradient that drives TEM. Although for the sake of clarity the T cell depicted here co-expresses the four chemokine receptors, not all CCR6 +(high) CCR2 + CD4 + T cells express CCR5 and/or CXCR3.

Article Snippet: Samples were incubated at room temperature for 2 h with primary mouse antibodies against human chemokines CCL2 (5 μg, Cat# MAB679) CCL5 (5 μg, Cat# MAB278), CCL7 (5 μg, Cat# MAB282), CCL8 (5 μg, Cat# MAB281), CCL20 (5 μg, Cat# AF360), CXCL9 (5 μg, Cat# MAB392) from R&D Systems or with mouse IgG 1 isotype control (5 μg, Cat# MAB002) or mouse IgG 2B isotype control (Cat# MAB004) from R&D Systems, or with anti-human GOLPH2 (2 μg, Cat# PA5-30622) or rabbit IgG isotype control from Invitrogen, or with anti-human vWF (2 μg, Cat# ab6994) from Abcam.

Techniques: Expressing, Binding Assay

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 1. Exposure to high altitude results in PH and increased secretion of inflammatory classical monocyte ligands from the lungs. (A) Schematic showing hypoxia exposure time course in wildtype mice. Duration of hypoxia exposure is directly proportional to (B) RVSP and RV hypertrophy as measured by Fulton Index (N=6-13/group). At 3 days of hypoxia, increased protein expression of classical monocyte ligands (C) CCL2 (N=6-11/group) and (D) CCL12 (N=6- 11/group), whereas significantly lower levels of nonclassical monocyte ligand (E) CX3CL1 (N=6/group) in the lungs. (F) Higher CCL2 gradient in lungs and in the (G) peripheral blood of wildtype mice following 3 days of hypoxia exposure (N=5/group). Data in all panels were obtained from female mice. Statistical analysis was conducted using ANOVA, followed by Tukey's post hoc test. *P<0.05, **P<0.01, ****P<0.0001. N=number of animals, mean±SD, CI=confidence interval.

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Expressing

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 4: Genetic and pharmacologic blockade of CCR2-CCL2 axis protects from hypoxic PH. (A) Schematic showing the BM reconstitution of Ccr2-/- and WT BM into lethally irradiated wildtype mice. Wildtype mice reconstituted with Ccr2-/- BM were protected from hypoxic PH by attenuated (B) RVSP (N=7-11/group) and (C) RV hypertrophy (N=7-11/group) as measured by Fulton Index, compared to wildtype mice that were reconstituted with wildtype BM. (D) Schematic showing pharmacological blockade of CCR2 ligands CCL2 or CCL7 using anti-CCL2 or anti-CCL7 neutralizing antibody treatment. Hypoxia exposed wildtype mice treated with CCL2 NAb but not CCL7 NAb showed lower (E) RVSP (N=6/group) and (F) RV hypertrophy (N=6/group). TSP-1 levels in (G) lungs (N=6/group) and (H) blood (N=6/group); and TGF-β1 levels in (I) lungs (N=6/group) and (J) blood (N=6/group) compared to wildtype mice treated with isotype control antibody. Data in all panels followed a normal distribution. ANOVA with the Tukey test was performed for multiple comparisons. Data were obtained from the female mice. mean ± SD

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Irradiation, Control

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 5: Resident IMs are a major source of CCL2 and recruited IMs are a major source of pathologic TSP-1 in hypoxic PH. (A) Flow cytometry analysis using Ccl2RFP-flox reporter mice showed a higher number of CCL2+ IMs (N=14/group; N=14/group, 9F and 5M in Nx; 8F and 6M in Hx), and (B) FOLR2+ IMs are a major source of CCL2 (N=14/group). (C) Hypoxia exposed wildtype mice following intracellular CCL2 staining by flow cytometry also showed a higher number of CCL2+ IMs (N=7/group, female mice). (D). IM subpopulation analysis using flow

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Flow Cytometry, Staining

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 8: DEX prophylaxis blunts CCL2 production by resident IMs and blocks the recruitment of TSP-1 producing CCR2+ IMs in hypoxia. (A) DEX prophylactically-treated, hypoxia-exposed Ccl2RFP-flox reporter mice exhibited a significant reduction in CCL2+ IMs, particularly in (B) CCL2RFP+ resident IMs (N=7/group). Additionally, (C) intracellular CCL2 flow cytometry analysis in DEX prophylactically-treated hypoxia-exposed wildtype mice revealed a

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Flow Cytometry