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Anti mac3

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Anti-Mac3 is a laboratory product used for the detection and quantification of Mac3, a protein commonly expressed in macrophages. It is designed to assist researchers in their investigations related to macrophage biology and associated research areas.

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Lab products found in correlation

Anti ki67, by Abcam (3 mentions) A2547, by Merck Group (3 mentions) Sirius red f3b, by Merck Group (2 mentions) Cm3050, by Leica (1 mentions) Anti cd8, by Merck Group (1 mentions) Sm α actin, by Merck Group (1 mentions) Image pro plus software, by Media Cybernetics (1 mentions) Anti cd4, by BD (1 mentions) Axiovision rel 4, by Zeiss (1 mentions) Masson s trichrome staining, by Merck Group (1 mentions) Anti cd31, by BD (1 mentions) Ab1893, by Abcam (1 mentions) Ab64693, by Abcam (1 mentions) Ab16667, by Abcam (1 mentions) Anti cd68 clone kp 1, by Agilent Technologies (1 mentions) A0452, by Agilent Technologies (1 mentions) Anti ym1, by R&D Systems (1 mentions) Anti cd45r, by BD (1 mentions) Ab15323, by Abcam (1 mentions) Ab124432, by Abcam (1 mentions)

Close spelling variants of this product

Anti-Mac3 antibody (10 citations) Anti-Mac3 clone M3/84 (2 citations)

18 protocols using anti mac3

1

Quantifying Atherosclerosis in LDLR-/- Mice

Cited 1 time
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To quantify atherosclerosis in LDLR−/− mice that were placed on HCD (Research Diets Inc., D12108C), aortic roots and aortic arch were embedded in OCT and frozen at −80 °C. Serial cryostat sections (6 µm) were prepared using tissue processor Leica CM3050. For lesion characterizations, the thoracic-abdominal aorta and aortic root were stained for ORO, macrophages (anti-Mac3, BD Pharmingen, 553322, 1:900) T cells (anti-CD4, BD Pharmingen, 553043, 1:90; anti-CD8, Chemicon, CBL1318, 1:100), MHC-positive cells (anti-MHCII, Novus Biologicals), and vascular smooth muscle cells (SM-α-actin, Sigma, F-3777, 1:500)73 (link),83 (link). The staining area was measured using Image-Pro Plus software, Media Cybernetics, and CD4+ and CD8+ cells were counted manually.
Simion V., Zhou H., Haemmig S., Pierce J.B., Mendes S., Tesmenitsky Y., Pérez-Cremades D., Lee J.F., Chen A.F., Ronda N., Papotti B., Marto J.A, & Feinberg M.W. (2020). A macrophage-specific lncRNA regulates apoptosis and atherosclerosis by tethering HuR in the nucleus. Nature Communications, 11, 6135.
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2

Histological Evaluation of Cardiac Remodeling

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Histological analysis was performed in 10 µm cross sections with a distance of 200 µm. The following antibodies were used to estimate angiogenesis, cell proliferation and macrophage infiltration: anti-CD31, anti-Mac 3 (BD Pharmingen, Franklin Lakes, USA), anti-Ki-67 (Abcam, Cambridge, UK), rabbit anti-rat and goat anti-rabbit secondary antibodies (VectorLabs, Burlingame, CA). TdT-mediated dUTP Nick-End Labeling (TUNEL) was performed using a kit system according to the manufacturer’s protocol (Promega, Madison, USA). Mean amount of positive signals per three sections were calculated as fold change relative to Sham operated animals. Collagen deposition and cardiomyocyte size were determined using Masson‘s trichrome staining (Sigma, Weelze, Germany) and wheat germ agglutinin (WGA) staining (Life Technologies, Darmstadt, Germany), respectively [30 (link)]. The area of fibrosis was determined in 4 randomly chosen sections with 4 randomly chosen areas per section, while blindly moving the microscope at 20× magnification. A grid was placed over the image, followed by counting the boxes that contained blue dye using ImageJ software. Cardiomyocyte size was determined in 3 randomly chosen sections per sample and the mean size of 30 cardiomyocytes was calculated using AxioVision Rel 4.6 (Carl Zeiss AG, Feldbach, Switzerland) at 40× magnification.
Koentges C., Pepin M.E., Müsse C., Pfeil K., Viteri Alvarez S.V., Hoppe N., Hoffmann M.M., Odening K.E., Sossalla S., Zirlik A., Hein L., Bode C., Wende A.R, & Bugger H. (2017). Gene Expression Analysis to Identify Mechanisms Underlying Heart Failure Susceptibility in Mice and Humans. Basic research in cardiology, 113(1), 8.
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3

Multiparametric Immunohistochemistry for Immune Profiling

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Antibodies included anti‐BrdU, ab1893 (Abcam, Cambridge, MA), heat‐induced epitope retrieval (HIER; pH 6.0), used at 1:500; anti‐Ki67, ab16667 (Abcam), HIER, 1:100; anti‐Mac‐3, #55322 (BD Pharmingen, San Diego, CA), HIER, 1:1000; anti‐CD3, A 0452 (DakoCytomation, Carpinteria, CA), HIER, 1:100; anti‐CD45R, #550286 (Pharmingen), HIER, 1:50; anti‐mannose receptor (MR), ab64693 (Abcam), HIER, 1:1000; anti‐YM1 (chitinase 3‐like 3), R&D Systems (Minneapolis, MN), HIER, 1:1000; anti‐iNOS (inducible nitric oxide synthase), ab15323, (Abcam), HIER, 1:100; anti‐arginase I (Arg I), BD Transduction, 1:100; anti‐FOXP3, FJK‐16 (eBioscience, San Diego, CA), HIER, 1:10; anti‐MPO (myeloperoxidase), 120‐15484 (Novus, Saint Charles, MO), HIER, 1:10; anti‐SMA (smooth muscle actin), A2547 (Sigma‐Aldrich), 1:200; anti‐CD68, clone KP1 (DAKO), HIER, 1:100; anti‐CD4, 14‐9766 clone 4SM95 (Affymetrix, Santa Clara, CA), HIER, 1:50; and anti‐CD8, 14‐0808 Clone 4SM15 (Affymetrix), HIER, 1:100. Isotype and preimmune negative controls were used during staining optimization and lacked background staining.
Lhoták Š., Gyulay G., Cutz J., Al‐Hashimi A., Trigatti B.L., Richards C.D., Igdoura S.A., Steinberg G.R., Bramson J., Ask K, & Austin R.C. (2016). Characterization of Proliferating Lesion‐Resident Cells During All Stages of Atherosclerotic Growth. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(8), e003945.
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4

Analyzing Vein Graft Lesions and Neoangiogenesis

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After euthanasia, vein graft segments were collected, fixed in 4% paraformaldehyde (PFA) for 24 h, dehydrated overnight in 60% isopropanol, and subsequently embedded in paraffin. Cross sections of vein graft segments were stained with hematoxylin and eosin to evaluate lumen and lesion area, plaque thickness, and percentage of vein stenosis. Neovessels were detected inside vein graft lesions via standard immunohistochemistry using anti-CD31 antibody (endothelial cells; ab124432, Abcam). Anti-TER-119 (550,565, BD Biosciences) was used to determine plaque hemorrhages and anti-α-smooth muscle actin (α-SMA) (A2547, Sigma-Aldrich) was used to determine vascular smooth muscle cell (VSMC) coverage of neovessels. Anti-MAC3 (550,292, Pharmingen), a Masson’s Trichrome stain and anti-vascular cell adhesion molecule-1 (VCAM-1) (ab134047, Abcam) were used to stain macrophages, collagen, and VCAM-1 positive ECs, respectively.
Perrotta P., de Vries M.R., Peeters B., Guns P.J., De Meyer G.R., Quax P.H, & Martinet W. (2021). PFKFB3 gene deletion in endothelial cells inhibits intraplaque angiogenesis and lesion formation in a murine model of venous bypass grafting. Angiogenesis, 25(1), 129-143.
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5

Immunostaining Protocol for Tissue Analysis

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The following antibodies were used for immunostaining: anti-AIF1 (1:500 dilution) from ThermoFisher (Waltham, MA), anti-GSDMD (1:500 dilution) from Santa Cruz (Dallas, TX), anti-MAC-3 (1:20 dilution) from BD Biosciences (San Jose, CA), anti-vWF (1:50 dilution) from Dako (Carpinteria, CA), anti-α-SMA (1:100 dilution) from Sigma (Saint Louis, MO), anti-Ki67 (1:100 dilution), anti-neutrophil elastase (1:4000 dilution) and anti-RBPMS (1:500 dilution) from Abcam (Cambridge, MA). AlexaFluor-594 labeled isolectin IB4, TUNEL assay, and all qRT-PCR primers were purchased from ThermoFisher.
Sonny S., Yuan H., Chen S., Duncan M.R., Chen P., Benny M., Young K., Park K.K., Schmidt A.F, & Wu S. (2023). GSDMD deficiency ameliorates hyperoxia-induced BPD and ROP in neonatal mice. Scientific Reports, 13, 143.
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6

Immunohistochemical Analysis of Ischemic Muscle

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Mice were euthanized 7 days after surgery. The ischaemic gastrocnemius muscles were excised, embedded in paraffin or TISSUE TEK OCT compound, and stored at −80 °C. Cross-sections were prepared for immunohistochemical analysis. Capillary density was determined by immunostaining with anti-PECAM-1 antibody. The number of PECAM-1+ vessels was quantified in 5 microscopic fields in each of 3 cross-sections from each tissue specimen using ImagePro Plus software. Adductor muscle cryostat sections were obtained, and incubated with primary antibodies anti-Mac-3 (1:500, Pharmingen) and α-SMA (1:100, Santa Cruz Biotech). The secondary antibodies used were goat ant-rat IgG AlexaFluor 488-conjugated antibody and goat ant-rabbit IgG AlexaFluor 568-conjugated antibody (Molecular Probes, Invitrogen). Images were captured by a fluorescence microscopy (Leica).
In some experiments, cross-sections were stained with hematoxylin-eosin (HE). Photomicrographs were taken using a microscope (Leica), the collateral diameters were measured in 5 randomly selected fields per section. The mean value of these measurements was taken as a single point for each animal.
Tan X., Yan K., Ren M., Chen N., Li Y., Deng X., Wang L., Li R., Luo M., Liu Y., Liu Y, & Wu J. (2016). Angiopoietin-2 impairs collateral artery growth associated with the suppression of the infiltration of macrophages in mouse hindlimb ischaemia. Journal of Translational Medicine, 14, 306.
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7

Quantifying Aortic Lesion Composition

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For quantification of collagen deposition, paraffin-embedded lesions of aortic root were stained with Sirius Red F3B (CI 35782; Sigma-Aldrich). For macrophage content, sections of paraffin-embedded or cryo-sections of aortic root lesions were stained with an anti–MAC-3 (BD Pharmingen) or anti-MOMA2 Ab (Bio-Rad), respectively, as described previously.13 (link),15 (link) For CXCL10 quantification, sections were stained with a rabbit anti-CXCL10 Ab (using the commercially available 10H11L3 clone that has been validated in paraffin-embedded spleen sections and in Western blot analysis of recombinant murine CXCL10 by the manufacturer; ThermoFisher). For caspase-3 quantification, we used an anticaspase-3 Ab (Cell Signaling) that specifically detects the cleaved form of caspase-3. Quantification was performed with computer-assisted image analysis using Adobe Photoshop Elements 6.0 and ImageJ software.
Tsiantoulas D., Sage A.P., Göderle L., Ozsvar-Kozma M., Murphy D., Porsch F., Pasterkamp G., Menche J., Schneider P., Mallat Z, & Binder C.J. (2018). B Cell–Activating Factor Neutralization Aggravates Atherosclerosis. Circulation, 138(20), 2263-2273.
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8

Quantifying Inflammatory Cell Infiltration

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Immunohistochemical studies were performed by immunoperoxidase methods using paraffin‐embedded tissue sections (6 mm thick). The sections were stained with haematoxylin and eosin to identify infarct area. The sections were incubated with primary anti‐myeloperoxidase (MPO; Abbiotec, UK), anti‐Mac3 (BD Bioscience), and anti‐CD3 (eBioscience) at 4°C overnight, followed by respective secondary HRP‐conjugated antibodies for 1 hour at room temperature. The positive cells were visualized with DAB, and nuclei were counterstained with hematoxylin. The numbers of MPO+ neutrophils, mouse CD107b (Mac3+) macrophages, and CD3+ T lymphocytes were assessed by counting the total cell numbers in the infarcted and border areas in 10 randomly chosen fields in each section.
Zhu R., Sun H., Yu K., Zhong Y., Shi H., Wei Y., Su X., Xu W., Luo Q., Zhang F., Zhu Z., Meng K., Zhao X., Liu Y., Mao Y., Cheng P., Mao X, & Zeng Q. (2016). Interleukin‐37 and Dendritic Cells Treated With Interleukin‐37 Plus Troponin I Ameliorate Cardiac Remodeling After Myocardial Infarction. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(12), e004406.
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9

Aortic Plaque Characterization with Histology

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4% PFA-fixed frozen aortic root sections with 5-μm-thickness were used for all studies. To assess plaque complexity, necrotic core and collagen content were visualized by hematoxylin and eosin (H&E, #HHS80 and #HT110180) and Masson-Goldner's trichrome staining (#100485, all purchased from Sigma-Aldrich, United States), which were performed according to the manufacturer's protocols. For immunofluorescent staining, slides were permeabilized with 0.5% tritonX-100 for 10 min, followed by blocking with 5% donkey serum (#D9663, Sigma-Aldrich, United States) in 0.5% tritonX-100 for 30 min at room temperature. Anti-Itga9 (#AF3827, 1:40, R&D systems, United States) was used to detect Itga9 expression, and normal goat IgG (#NI02-100UG, 1:40, EMD Millipore, United States) was used as isotype control. Both antibodies were then visualized with donkey anti-goat Alexa594 (#A11058, 1:400, Invitrogen, United States). Anti-Mac3 (#550292, 1:100, BD Biosciences, United States), anti-SMα-actin-cy3 (#C6198, 1:500, Sigma-Aldrich, United States) were used, and then visualized with donkey anti-rat Alexa488 (#A21208, 1:400, Invitrogen, United States) was used. Nuclear counterstaining and slide mounting were performed with Prolong Diamond Antifade with DAPI (#P36862, Invitrogen, United States).
Jung I.H, & Stitziel N.O. (2024). Integrin α9β1 deficiency does not impact the development of atherosclerosis in mice. Heliyon, 10(4), e25760.
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10

Immunohistochemical Analysis of Angiogenic Factors

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As indicated, antibodies included rat monoclonal anti-mouse CD31 (1:50, BD Pharmingen); mouse monoclonal and Cy3-conjugated anti-mouse smooth muscle a-actin SMA (1:200, Sigma); anti- phospho-histone H3 (PH3, Cell Signaling Technology), anti-Flk1 (1:100, Cell Signaling Technology), anti-Flt-1 (1:50, Cell Signaling Technology), anti-mouse F4/80 (1:100, eBioscience); anti-MAC3 (1:100, BD Pharmingen), anti-mouse VEGF (1:50 Neomarkers Inc). Secondary antibodies included Alexa Fluor® 594 labeled anti-rat or anti-rabbit IgG (1:1000, Jackson Immuno Res) and Alexa Fluor 488-conjugated goat anti-rat IgG (H+L) (Molecular Probes). Growth factors include human recombinant vascular endothelial growth factor (VEGF165, R&D Systems), human recombinant fibroblast growth factor (FGF2) (R&D Systems), epidermal growth factor (EGF) (Cell Signaling Technology), insulin-like growth factor 1 (IGF-1) (Cell Signaling Technology), stem cell factor (SCF) and interleukin 3 (IL-3) (R&D Systems).
Han Y., Tao J., Gomer A, & Ramirez-Bergeron D.L. (2014). Loss of endothelial-ARNT in adult mice contributes to dampened circulating proangiogenic cells and delayed wound healing. Vascular medicine (London, England), 19(6), 429-441.
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