Antibodies directed against mouse CD31 (PECAM-1) (affinity purified SL-4), raised in rabbit and purified as described elsewhere 20 (link). Rat monoclonal antibodies against mouse CD31 [clone: MEC13.3] (Cat No. 550274), CD45 [30-F11] (Cat No. 550539), CD11b [M1/70] (Cat No. 550282), Mac3 [M3/84] (Cat No. 550292), and VE-cadherin (VEC) [11D4.1] (Cat No. 550548) were purchased from BD Biosciences (San Jose, CA, USA). Rabbit polyclonal antibodies against Ki67 (Cat No. ab15580) were purchased from Abcam (Cambridge, MA, USA). Rabbit monoclonal antibodies against Survivin [71G4B7] (Cat No. #2808), YAP [D24E4] (Cat No. #8418), and phospho-YAP (Ser127) [D9W2I] (Cat No. #13008) (P-YAP) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). A mouse monoclonal antibody against glucose transporter 1 (GLUT-1) [SPM498] (Cat No. ab40084) was purchased from Abcam (Cambridge, MA, USA). Mouse monoclonal antibodies against human CD31 [JC70A] (Cat No. M0823), a CD45 [2B11+PD7/26] (Cat No. M0701) and a monoclonal mouse anti human CD68 were purchased from DAKO. Rabbit polyclonal antibodies against human Ajuba (Cat No. HPA006171) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). A rabbit polyclonal against human GLUT1 was purchased from Cell Marque (Rocklin, CA).
Monoclonal mouse anti human cd68
Manufactured by Agilent Technologies
Sourced in Denmark
The Monoclonal mouse anti human CD68 is a laboratory reagent used to detect the presence of the CD68 protein, which is a marker for macrophages and monocytes in human tissue samples. This product is intended for research use only.
Automatically generated - may contain errors
Lab products found in correlation
Cd45 2b11 pd7 26, by Agilent Technologies (1 mentions)
Cd11b m1 70, by BD (1 mentions)
Cd45 30 f11, by BD (1 mentions)
Mac3 m3 84, by BD (1 mentions)
Yap d24e4, by Cell Signaling Technology (1 mentions)
Spm498, by Abcam (1 mentions)
Monoclonal mouse anti human cd45, by Agilent Technologies (1 mentions)
Monoclonal mouse antihuman mast cell tryptase, by Agilent Technologies (1 mentions)
Colchicine, by Merck Group (1 mentions)
Bond polymer refine detection system, by Leica (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Bz 9000 microscope, by Keyence (1 mentions)
Dab substrate kit, by Nichirei Biosciences (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti alpha smooth muscle actin α sma, by Abcam (1 mentions)
6 protocols using monoclonal mouse anti human cd68
1
Antibody Characterization for Vascular Research
2
Immunohistochemical Analysis of Colon Tissue
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The tissue samples taken from the colon during postmortem laparotomy were fixed overnight in 4% buffered paraformaldehyde, embedded in paraffin, and then cut into 5-µm-thick sections. The sections were deparaffinized and then stained with hematoxylin and eosin (H&E) or else immunostained using the ultraView Universal DAB Detection kit (v1.02.0018) and the BenchMark Ultra IHC/ISH staining module (both from Venata Medical Systems, Basel, Switzerland). For immunostaining, the sections were incubated with one of the primary antibodies for 32 min at 37°C. The primary antibodies used were monoclonal mouse antihuman CD45 (code no. M0701), monoclonal mouse anti-human CD47 (code no. I5647), monoclonal mouse antihuman CD68 (code no. M0814) and monoclonal mouse antihuman mast cell tryptase (code no. M7052) (all from Dako, Glostrup, Denmark). CD45 is considered as a common leukocyte antigen and is expressed exclusively on cells of the hematopoietic system and their progenitors. CD57 is expressed by subsets of natural killer cells and CD8+ lymphocytes, and by a small proportion of CD4+/CD45R0+ T lymphocytes. CD68 labels human monocytes, macrophages and myeloid cells. Human mast cell tryptases comprise a family of trypsin-like neutral serine proteases that are expressed predominantly in mast cells.
3
Hepatic Cytokine and Macrophage Quantification
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The liver tissue blocks were obtained from surgery for HBV-HCC (n = 30), liver biopsy for CHB liver (n = 5) or HBV-cirrhosis liver (n = 5) and the off cuts of liver transport donors for HCs (n = 5). HCs did not present liver disease/HBsAg negative/negative image in CT or MRI. Hepatic IL-34, MCSF and CD68 were determined using immunohistochemistry (IHC), using 3,3′-diaminobenzidine (DAB) color development. The primary antibodies were polyclonal rabbit anti-human IL-34 (bs-18170R, Beijing Biosynthesis Biotechnology, China), polyclonal rabbit anti-human MCSF (Abcam, Cambridge, UK) and monoclonal mouse anti-human CD68 (Dako, Copenhagen, Denmark). The sheep anti-rabbit conjugated HRP secondary antibody (Beijing Sequoia Jinqiao Biological Technology) was used. The specific target(s) was visualized with DAB detection kit and counterstained with hematoxylin. The IHC was repeated twice. Negative control was applied in each labeling for every primary rabbit negative control. Intra-hepatic IL-34 or MCSF is localized in the cytoplasm of hepatocytes, which has been well documented in our previous publications [28 (link)]. IHC was quantified using a computer-assisted genuine color image analysis system (ImagePro-plus 9.0) for hepatic IL-34, MCSF or CD68, as described previously [28 (link), 29 (link)].
4
Evaluating RS and DPZ Neuroprotective Effects
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RS (pale yellow powder, 99.9% pure form) and colchicine were received from Bengaluru, India (Sigma-Aldrich, catalog number 9754). The mouse monoclonal anti-glial fibrillary acidic protein (GFAP) and mouse monoclonal anti-human CD68 were procured from Dako, Denmark, and Produktionsverg, Denmark. Other chemicals were obtained from HPLC, Sigma (St. Louis, USA). The RS dosage was decided as per Sharma and Gupta (2002 (link)) and Wiciński et al. (2020 (link)). The DPZ dose ranges between 0.375 and 0.75 mg/kg/day as per Hernandez et al. (2006 (link)). In this study, the dosage of DPZ (1 mg/kg/day) and RS (10 mg and 20 mg/kg/day) was considered as per our previous research (Rao et al. 2021 (link)).
5
Adipocyte Density and Macrophage Dynamics in Bariatric Surgery
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We aimed to collect two fat tissue samples from all subjects, one at surgery and the second one a year after bariatric surgery (surgery group) or before and after receiving conservative treatment for 1 year (conservative group). Ultimately, after dropouts, refusals, and additional data loss, 39 (data missing for 21) and 43 (data missing for 19) paired samples were analyzed in the surgery and conservative groups, respectively. Missing data were omitted in analysis. An SAT biopsy specimen (circa 1 cm3) from the periumbilical area was obtained during the surgery or clinic appointment under local anesthesia. Tissue biopsy specimens were fixed in formalin and embedded in paraffin. The specimens were sectioned at 5 μm. One section was stained with hematoxylin and eosin (H&E) to assess adipocyte density. For CD68, immunohistochemistry was performed with BOND Polymer Refine Detection System (DS9800; Leica Biosystems, Buffalo Grove, Illinois) with 3,3′‐diaminobenzidine as a chromogen. Mouse monoclonal anti‐human CD68 (1:200; Dako; Agilent, Santa Clara, California) was used for 30 minutes with 20 minutes of Tris‐EDTA pretreatment.
6
Antibody-based Adipocyte Characterization
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Rabbit polyclonal antibodies targeting PLIN1 or PLIN2 were generated as described previously27)
. Mouse monoclonal anti-human CD68 (1:100, Dako Cytomation, Glostrup, Denmark), mouse monoclonal anti-CD11c (1:100, Dako), mouse monoclonal anti-human CD163 (1:100, Leica Biosystems, Wetzlar, Germany), and anti-alpha smooth muscle actin (α-SMA) (1:100, Abcam, Cambridge, UK) antibodies were purchased. Histological specimens and cells were blocked with 10% goat non-immune serum and were then incubated with a primary antibody overnight at 4℃. They were then stained using a DAB substrate kit (Nichirei Bioscience, Tokyo, Japan). For immunofluorescence, cells were cultured on coverslips placed in dishes and then incubated with fluorescently-labeled antibodies and Alexa Fluor 594-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific, Waltham, MA, US) for 60 min, BODIPY lipid probes for triglyceride (Molecular Probes, Eugene, OR, US) for 60 min, and DAPI (Thermo Fisher Scientific). Images were acquired using a BZ-9000 microscope (Keyence, Osaka, Japan).
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Rabbit polyclonal antibodies targeting PLIN1 or PLIN2 were generated as described previously27)
. Mouse monoclonal anti-human CD68 (1:100, Dako Cytomation, Glostrup, Denmark), mouse monoclonal anti-CD11c (1:100, Dako), mouse monoclonal anti-human CD163 (1:100, Leica Biosystems, Wetzlar, Germany), and anti-alpha smooth muscle actin (α-SMA) (1:100, Abcam, Cambridge, UK) antibodies were purchased. Histological specimens and cells were blocked with 10% goat non-immune serum and were then incubated with a primary antibody overnight at 4℃. They were then stained using a DAB substrate kit (Nichirei Bioscience, Tokyo, Japan). For immunofluorescence, cells were cultured on coverslips placed in dishes and then incubated with fluorescently-labeled antibodies and Alexa Fluor 594-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific, Waltham, MA, US) for 60 min, BODIPY lipid probes for triglyceride (Molecular Probes, Eugene, OR, US) for 60 min, and DAPI (Thermo Fisher Scientific). Images were acquired using a BZ-9000 microscope (Keyence, Osaka, Japan).
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