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: Biomedicines

Article Title: Increased Myocardial MAO-A, Atrogin-1, and IL-1β Expression in Transgenic Mice with Pancreatic Carcinoma—Benefit of MAO-A Inhibition for Cardiac Cachexia

doi: 10.3390/biomedicines12092009

Figure Lengend Snippet: QuantiTect primer assays used for qPCR.

Article Snippet: Transversal or longitudinal cryo-sections of the ventricular left myocardium were fixed by 5% paraformaldehyde (PFA) for 10 min, blocked with 1% normal swine serum (NSS) at 37 °C, and incubated with primary rabbit anti-mouse polyclonal antibodies against atrogin-1 (FBXO32, MAFbx, 1:200, ab74023, Abcam, Cambridge, UK), against IL-1β (1:50, ab9722, Abcam, Cambridge, UK), TNF (1:100, ab6671, Abcam, Cambridge, UK) or against COX2 (1:200, ab15191, Abcam, Cambridge, UK), and primary rat anti-mouse primary monoclonal antibodies against CD68 (1:50, MCA1957, AbD Serotec, Kidlington, UK).

Techniques: Amplification

Journal: Biomedicines

Article Title: Increased Myocardial MAO-A, Atrogin-1, and IL-1β Expression in Transgenic Mice with Pancreatic Carcinoma—Benefit of MAO-A Inhibition for Cardiac Cachexia

doi: 10.3390/biomedicines12092009

Figure Lengend Snippet: Expression of pro-atrophic, -inflammatory, -angiogenic, and -apoptotic transcripts in left myocardium of WT-HH, CA, and CA-HH mice relative to the group of untreated WT mice.

Article Snippet: Transversal or longitudinal cryo-sections of the ventricular left myocardium were fixed by 5% paraformaldehyde (PFA) for 10 min, blocked with 1% normal swine serum (NSS) at 37 °C, and incubated with primary rabbit anti-mouse polyclonal antibodies against atrogin-1 (FBXO32, MAFbx, 1:200, ab74023, Abcam, Cambridge, UK), against IL-1β (1:50, ab9722, Abcam, Cambridge, UK), TNF (1:100, ab6671, Abcam, Cambridge, UK) or against COX2 (1:200, ab15191, Abcam, Cambridge, UK), and primary rat anti-mouse primary monoclonal antibodies against CD68 (1:50, MCA1957, AbD Serotec, Kidlington, UK).

Techniques: Expressing

Journal: Biomedicines

Article Title: Increased Myocardial MAO-A, Atrogin-1, and IL-1β Expression in Transgenic Mice with Pancreatic Carcinoma—Benefit of MAO-A Inhibition for Cardiac Cachexia

doi: 10.3390/biomedicines12092009

Figure Lengend Snippet: CD68+ cells in the left ventricular myocardium of WT, WT-HH, CA, and CA-HH mice. ( a ) Percentage CD68+ cells. Data are given as mean ± SEM. Two-factorial ANOVA detected no significant impact of factor CA or factor HH treatment; however, revealed a significant interaction ( p ≤ 0.05) between these factors within the total study population. * p < 0.05 CA vs. WT. # p < 0.05 WT-HH vs. WT. ( b ) Representative photos of left myocardial cross-sections of WT, WT-HH, CA, and CA-HH mice showing CD68+ cells (arrows). ( c ) X-fold expression of CD68 transcripts in WT-HH, CA, and CA-HH relative to WT mice as assessed in pooled samples.

Article Snippet: Transversal or longitudinal cryo-sections of the ventricular left myocardium were fixed by 5% paraformaldehyde (PFA) for 10 min, blocked with 1% normal swine serum (NSS) at 37 °C, and incubated with primary rabbit anti-mouse polyclonal antibodies against atrogin-1 (FBXO32, MAFbx, 1:200, ab74023, Abcam, Cambridge, UK), against IL-1β (1:50, ab9722, Abcam, Cambridge, UK), TNF (1:100, ab6671, Abcam, Cambridge, UK) or against COX2 (1:200, ab15191, Abcam, Cambridge, UK), and primary rat anti-mouse primary monoclonal antibodies against CD68 (1:50, MCA1957, AbD Serotec, Kidlington, UK).

Techniques: Expressing

Journal: British journal of pharmacology

Article Title: Direct vagus nerve stimulation: A new tool to control allergic airway inflammation through α7 nicotinic acetylcholine receptor.

doi: 10.1111/bph.16334

Figure Lengend Snippet: FIGURE 4 Vagus nerve stimulation (VNS) 0.1 reduces the level of macrophages and goblet cells expressing α7nAChR. (a) Lung sections were stained for α7nAChR (red), CD68 (macrophages, green), and DNA nuclear stain (DAPI, blue). Scale: 50 μm. Macrophages and goblet cells are shown with higher magnification. Examples of double-staining are identified with white arrows. (b) Quantification of fluorescent intensity of α7nAChR in alveolar and peribronchial area. Values are shown as means ± SEM of five animals. *Significant difference from house dust mite (HDM): *P < 0.05. #Significant difference between VNS 0.1 and VNS 0.1 + α-bungarotoxin (α-BGTX): #P < 0.05.

Article Snippet: Briefly, tissue sections were incubated in 0.2% bovine serum albumin (BSA) blocking solution, followed by incubation over- night at 4 C of (i) anti-ChAT rabbit monoclonal primary antibody (EPR 16590, dilution 1:300; Abcam, Cambridge, UK, Cat# ab178850, RRID:AB_2721842) and Cy3-conjugated antirabbit secondary anti- body (dilution 1:300, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA, Cat# 611-164-215, RRID:AB_2922862) to detect ChAT, (ii) anti-α7nAChR rabbit monoclonal primary antibody (dilution 1:200; Alomone Labs, Jerusalem, Israel, Cat# ANC-007-FR, RRID:AB_2756676) and Cy3-conjugated antirabbit secondary anti- body (dilution 1:300, Jackson ImmunoResearch Laboratories, Cat# 611-164-215, RRID:AB_2922862) to detect α7nAChR, (iii) anti-CD68 rat monoclonal primary antibody (dilution 1:300; BIORAD, Hercules, CA, Cat# MCA1957, RRID:AB_322219) and Cy3-conjugated antirat secondary antibody (dilution 1:300, Jackson ImmunoResearch Laboratories, Cat# 611-164-215, RRID:AB_2922862) to detect macrophages.

Techniques: Expressing, Staining, Double Staining

Journal: Neuron

Article Title: Reorganization of adolescent prefrontal cortex circuitry is required for mouse cognitive maturation

doi: 10.1016/j.neuron.2023.10.024

Figure Lengend Snippet: Dendritic arborization complexity and spine density of prefrontal layer 2/3 pyramidal neurons throughout late development (A) Characteristic photographs of tDimer2-expressing L2/3 PYRs within the PL displayed together with average heatmaps of all overlayed dendrites for each age group. (B) Dendritic intersections within a 200 μm radius from the soma center of prelimbic L2/3 PYRs (n = 12 mice, 71 neurons). Color-striped bars indicate significant difference. (C) Violin plots displaying the total branch length (left) and number of branches (right) of the cells analyzed in (B). (D) Violin plots displaying spine densities of basal (i.e., originating from the soma; left, n = 17 mice, 60 neurons, 358 dendrites) and distal (i.e., originating from the apex/apical dendrite; right, n = 17 mice, 60 neurons, 251 dendrites) dendrites of L2/3 PYRs within the PL. (E) Characteristic photographs of basal dendrites from each age group. For line plots in (B), data are presented as mean ± SEM. Data of violin plots are presented as median with 25 th and 75 th percentile. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also Figure S6 and Table S1 for detailed statistics. (F) Characteristic photographs of Iba1 immunostainings in the PL of each age group. (G) Violin plot displaying the densities of Iba1-positive cells within the PL (n = 25 mice, 800 stacks). (H) Violin plots displaying roundness, cell spread, cell area, and cell perimeter for Iba1-positive cells within the PL (n = 25 mice, 26,707 cells). (I) Characteristic reconstructions of processed raw images of Iba1-positive cells (green) containing CD68- (blue) and PSD95- (red) positive puncta of L2/3 PYRs from each age group. Pink corresponds to an overlay of all three colors. (J) Bar diagrams displaying the percentage of microglia with colocalized puncta (n = 17 mice, 625 cells). (K) Violin plots displaying volume of vesicles (left) and inclusions (right), quantified on colocalized Iba1-, CD68- or Iba1-, CD68-, and PSD95-positive pixel, respectively (n = 17 mice, 625 cells). (L) Same as (K) for the number of inclusions. (M) Violin plot displaying the phagocytic ratio (i.e., MI of colocalized PSD95 vs. solely CD68 puncta, n = 17 mice, 625 cells). For line plots in (B), data are presented as mean ± SEM. For (K) and (L), data were normalized to Iba1 volume. Data of violin plots are presented as median with 25 th and 75 th percentile. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also Table S1 for detailed statistics.

Article Snippet: Rat monoclonal primary antibody against mouse CD68 , Bio-Rad , Cat# MCA1957GA, RRID: AB_324217.

Techniques: Expressing

Journal: Neuron

Article Title: Reorganization of adolescent prefrontal cortex circuitry is required for mouse cognitive maturation

doi: 10.1016/j.neuron.2023.10.024

Figure Lengend Snippet: Adult cognitive abilities and prefrontal morphology after microglia manipulation during adolescence (A) Left, color-coded heatmaps of the time spent at different locations within the open-field arena by Control I (left) and PLX I (right) P56–P60 mice. Right, violin plots displaying relative time spent in the center (given as %, left) and relative resting time (given as %, right) in the open-field arena (n = 41 mice). (B) Violin plots displaying grooming duration (left) and frequency (right) in the open-field arena (n = 41 mice). (C) Left, color-coded heatmaps of the time spent at different locations within the eight-arm maze by Control I (left) and PLX I (right) P56–P60 mice during all trials. Right, line plot displaying the number of working memory (left) and reference (right) errors for all 12 trials. Violin plots displaying the normalized slope of the fitted working memory (left) and reference memory (right) performance of individual mice (n = 14 mice). Color-striped bars indicate significant difference. (D) Left, schematic of the experimental protocol during odor discrimination. Right, violin plots displaying the number of trials (left) and average trial latency (right) required by each mouse to reach criterium (n = 27 mice). (E) Left, schematic of the experimental protocol during odor reversal. Right, violin plots displaying the number of trials (left), average trial latency (middle), and total errors (given as %, right) required by each mouse to reach criterium (n = 27 mice). (F) Characteristic photographs of tDimer2-expressing L2/3 PYRs within the PL of Control I (left) and PLX I (right) P56–P60 mice. (G) Left, dendritic intersections within a 200 μm radius from the soma center of prelimbic L2/3 PYRs (n = 6 mice, 37 neurons). Color-striped bars indicate significant difference. Right, violin plots displaying the total branch length (left) and the number of branches (right) of the cells analyzed left. (H) Violin plots displaying spine densities of basal (i.e., originating from the soma; left, n = 6 mice, 24 neurons, dendrites) and distal (i.e., originating from the apex/apical dendrite; right, n = 6 mice, 24 neurons, dendrites) dendrites of L2/3 PYRs within the PL. (I) Violin plots displaying the densities (left, n = 6 mice, 200 stacks) and roundness (right, n = 6 mice, 5504 cells) of Iba1-positive cells within the PL. (J) Characteristic reconstructions of processed raw images of Iba1-positive cells (green) containing CD68- (blue) and PSD95- (red) positive puncta of L2/3 PYRs from Control I (left) and PLX I (right) P58–P60 mice. Pink corresponds to an overlay of all three colors. (K) Left, violin plots displaying volume of inclusions (left) and the number of inclusions (right), quantified on colocalized Iba1-, CD68-, and PSD95-positive pixel (n = 6 mice, 270 cells). Right, bar diagrams displaying the percentage of microglia with colocalized puncta. For line plots in (C) and (G), data are presented as mean ± SEM. Data of violin plots are presented as median with 25 th and 75 th percentiles. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also Figures S7 and as well as Table S1 for detailed statistics.

Article Snippet: Rat monoclonal primary antibody against mouse CD68 , Bio-Rad , Cat# MCA1957GA, RRID: AB_324217.

Techniques: Control, Expressing

Journal: Neuron

Article Title: Reorganization of adolescent prefrontal cortex circuitry is required for mouse cognitive maturation

doi: 10.1016/j.neuron.2023.10.024

Figure Lengend Snippet: Long-lasting functional, behavioral, and morphological consequences of microglia manipulation during adolescence (A) Schematic of the experimental procedure. (B) Left, power spectra of LFP activity recorded in the PL of P98–P102 Control II and PLX II mice. Right, scatterplot of peak frequencies as a function of peak amplitudes for 1–12 and 12–100 Hz bands, respectively (n = 11 mice, 22 recordings). (C) Same as (B) for LFP activity recorded in the S1 (n = 11 mice, 22 recordings). (D) Top, color-coded heatmaps of the time spent at different locations within the open-field arena by Control II (left) and PLX II (right) P98–P102 mice. Bottom, violin plots displaying the relative time spent in the center (given as %, left), relative resting time (given as % middle), and grooming frequency (right) in the open-field arena (n = 24 mice). (E) Left, schematic of the experimental protocol of spontaneous alternations monitoring. Right, violin plots displaying the number of entries (left) and percentage of alternations (right) in the Y-maze (n = 24 mice). (F) Left, schematic of the experimental protocol during odor discrimination. Right, violin plots displaying the number of trials (left) and average trial latency (right) required by each mouse to reach criterium (n = 24 mice). (G) Left, schematic of the experimental protocol during odor reversal. Right, violin plots displaying the number of trials (left), average trial latency (middle), and total errors (given as %, right) required by each mouse to reach criterium (n = 24 mice). (H) Characteristic photographs of tDimer2-expressing L2/3 PYRs within the PL of Control II (left) and PLX II (right) P98–P102 mice. (I) Left, dendritic intersections within a 200 μm radius from the soma center of prelimbic L2/3 PYRs (n = 6 mice, 36 neurons). Color-striped bars indicate significant difference. Right, violin plots displaying the total branch length (left) and number of branches (right) of the cells analyzed left. (J) Violin plots displaying spine densities of basal (i.e., originating from the soma; left, n = 6 mice, 24 neurons, 121 dendrites) and distal (i.e., originating from the apex/apical dendrite; right, n = 6 mice, 24 neurons, 101 dendrites) dendrites of L2/3 PYRs within the PL. (K) Violin plots displaying the densities (left, n = 6 mice, 200 stacks) and roundness (right, n = 6 mice, 6,149 cells) of Iba1-positive cells within the PL. (L) Characteristic reconstructions of processed raw images of Iba1-positive cells (green) containing CD68- (blue) and PSD95- (red) positive puncta of L2/3 PYRs from Control II (left) and PLX II (right) P98–P102 mice. Pink corresponds to an overlay of all three colors. (M) Left, violin plots displaying volume of inclusions (left) and the number of inclusions (right), quantified on colocalized Iba1-, CD68-, and PSD95-positive pixel (n = 6 mice, 252 cells). Right, bar diagrams displaying the percentage of microglia with colocalized puncta. For line plots in (B), (C), and (I), data are presented as mean ± SEM. Data of violin plots are presented as median with 25 th and 75 th percentiles. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also Table S1 for detailed statistics.

Article Snippet: Rat monoclonal primary antibody against mouse CD68 , Bio-Rad , Cat# MCA1957GA, RRID: AB_324217.

Techniques: Functional Assay, Activity Assay, Control, Expressing

Journal: Neuron

Article Title: Reorganization of adolescent prefrontal cortex circuitry is required for mouse cognitive maturation

doi: 10.1016/j.neuron.2023.10.024

Figure Lengend Snippet:

Article Snippet: Rat monoclonal primary antibody against mouse CD68 , Bio-Rad , Cat# MCA1957GA, RRID: AB_324217.

Techniques: Virus, Recombinant, Software, Imaging, Electroporation, Microscopy

Journal: Neurobiology of Pain

Article Title: Low-intensity pulsed ultrasound phonophoresis with diclofenac alleviated inflammation and pain via downregulation of M1 macrophages in rats with carrageenan-induced knee joint arthritis

doi: 10.1016/j.ynpai.2023.100148

Figure Lengend Snippet: Immunohistochemical staining for macrophages in the knee joint synovium. Representative photomicrographs of CD68, CD11c, CD206 immunohistochemistry in the knee joint synovium are shown (A). Arrowheads indicate each positive cell. The number of CD68 (B), CD11c (C), CD206 (D) positive cells /mm 2 in the knee joint synovium was calculated. The data are presented as mean ± standard deviation. * p < 0.05 significantly different from the control group. # p < 0.05 significantly different from the arthritis group. † p < 0.05 significantly different from the diclofenac group. ‡ p < 0.05 significantly different from the LIPUS group. Scale bar = 50 µm.

Article Snippet: Sections were blocked with 1 % bovine serum albumin in PBS for 60 min and incubated overnight at 20 °C with mouse monoclonal anti-CD68 primary antibody (1:3000 mouse, BIO-RAD), rabbit polyclonal anti-CD11c antibody (1:2000 rabbit, Funakoshi) and rabbit polyclonal anti-CD206 antibody (1:2000 rabbit, Abcom) as markers for the total, M1, and M2 macrophages, respectively.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Standard Deviation, Control

Journal: International journal of molecular sciences

Article Title: Long-Term Pioglitazone Treatment Has No Significant Impact on Microglial Activation and Tau Pathology in P301S Mice.

doi: 10.3390/ijms241210106

Figure Lengend Snippet: Figure 3. (a,b) Microglial activation (area%) in the cortex and the brainstem of P301S mice at 8.2 months of age treated with pioglitazone (P301S Pio) or placebo (P301S Placebo) as quantified by analyzing triplicates of immunohistochemical stainings with Iba1 (a) and CD68 (b). (c,d) Rep- resentative orthogonal projections of immunohistochemical stainings of P301S mice treated with pioglitazone (P301S Pio) or placebo (P301S Placebo) in the cortex (c) and the brainstem (d).

Article Snippet: To assess the degree of activation of microglia, paraformaldehyde-fixed 50 μm thick sagittal brain sections were incubated overnight at 4 ◦C in PBS with 5% normal goat serum and 0.5% Triton X-100 containing guinea pig monoclonal anti-Iba1 primary antibody (1:500, Synaptic Systems GmbH, Göttingen, Germany, 234308) and rat monoclonal anti-CD68 primary antibody (1:500, FA-11, Bio-Rad Laboratories Inc., Hercules, CA, USA, MCA1957).

Techniques: Activation Assay, Immunohistochemical staining