primary rat anti cd68 monoclonal antibody  (Bio-Rad)

Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—5 mg/kg). A ) Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B ) IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported, scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm. C) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, ** p ≤ 0.01. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-ACT, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E ) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—3 mg/kg). A Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm C ) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-actin, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: ( A ) Representative images of hematoxylin-eosin (H&E) staining and immunohistochemistry for CD68-positive macrophages in rat hearts collected 21 days after the first immunization. Rats were administered EC2319 or saline throughout the induction of myocarditis. ( B ) Quantification of CD68-positive macrophages in myocardial tissue on Day 21 of the prevention sub-study.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Staining, Immunohistochemistry, Saline

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: ( A ) Representative [ 18 F]FOL autoradiographs and corresponding hematoxylin and eosin (H&E) and immunohistochemical stainings of myocarditis rat hearts post-treatment. ( B ) Quantification of CD68-positive macrophages and ( C ) correlation with myocardial [ 18 F]FOL uptake on Day 26. Spearman correlation coefficient was used to analyze linear correlations between CD68-positive area and myocardial [ 18 F]FOL uptake on Day 26. ( D ) Quantifications of M1-polarized macrophages (iNOS-positive), ( E ) M2-polarized macrophages (MRC-1-positive), and ( F ) M1/M2 macrophage ratios in inflammatory lesions based on immunostaining. ( G ) Quantification of fibrotic area using Masson’s trichrome staining. n = 6-8 per group for B-C; n = 3 per group for D-F; n = 5-7 per group for G. Statistical comparisons of normally distributed data were performed using one-way ANOVA followed by post hoc tests: Tukey’s test (for homogeneous variances) and Games–Howell test (for non-homogeneous variances) to compare different treatments. Non-normally distributed continuous data are presented as median (interquartile range, IQR) and compared using the Kruskal–Wallis test, followed by pairwise comparisons with Bonferroni-adjusted p -values.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Immunohistochemical staining, Immunostaining, Staining

Journal: bioRxiv

Article Title: Targeting Macrophage Folate Receptor-β for Diagnosis and Treatment of Autoimmune Myocarditis

doi: 10.1101/2025.11.02.686077

Figure Lengend Snippet: There was strong co-localization of CD68 and iNOS, a marker of pro-inflammatory M1 macrophages, whereas there was only partial co-localization of CD68 and MRC-1, a marker of anti-inflammatory M2 macrophages. Number of iNOS-positive macrophages is markedly lower after treatment with EC2319 than saline or Etanercept, consistent with a shift toward an anti-inflammatory macrophage profile.

Article Snippet: For immunohistochemistry, adjacent serial 8-μm cryosections were stained with monoclonal mouse anti-rat CD68 antibody (1:1000, MCA341GA, Bio-Rad) to identify macrophages.

Techniques: Marker, Saline