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Cd68 fa 11

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CD68 (FA-11) is a monoclonal antibody that recognizes the CD68 antigen, a glycoprotein expressed primarily by macrophages and monocytes. The CD68 antigen is a lysosomal-associated protein that is widely used as a marker for the identification and characterization of macrophages and monocytes in various tissues and cell populations.

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Lab products found in correlation

Facsdiva, by BD (3 mentions) Ly6g 1a8, by BD (3 mentions) Leucoperm, by Bio-Rad (2 mentions) F4 80 bm8, by Thermo Fisher Scientific (2 mentions) Mertk 108928, by R&D Systems (2 mentions) Cd14 sa14 2, by BioLegend (2 mentions) Facscanto, by BD (2 mentions) Cellquest, by BD (2 mentions) Facs caliber, by BD (2 mentions) Siglecf e50 2440, by BD (2 mentions) Ly6g 1a8, by BioLegend (2 mentions) Cd103 2e7, by BioLegend (2 mentions) Mmp 9 af909, by R&D Systems (2 mentions) Prolong gold mountant, by Thermo Fisher Scientific (1 mentions) Moma 1, by Abcam (1 mentions) Cd138 281 2, by BioLegend (1 mentions) B220 ra3 6b2, by BioLegend (1 mentions) Golgiplug, by BD (1 mentions) Cytofix cytoperm, by BD (1 mentions) Monensin, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

FA-11 (10 citations) Anti-CD68 (clone FA-11, (3 citations) Anti-CD68 mAb (clone FA-11, (2 citations) CD68 (clone FA-11) (2 citations) Biotin-conjugated anti-CD68 Ab (FA-11, (2 citations) Monoclonal rat anti-CD68 antibody (Clone FA-11) (2 citations)

14 protocols using cd68 fa 11

1

Purification and Immunophenotyping of BAL Cells

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BAL cells were purified by centrifugation on Percoll to remove surfactant and debris 31 (link). BAL cells or BMDMs were immunostained to detect CD115 (AFS98), F4/80 (BM8) (eBioscience), CD3 (145-2C11), CD11b (M1/70), CD11c HL3), CD16/32 (2.4G2), CD19 (1D3), CD64 (X54-5/7.1.1), CD131 (JORO50), Ly6G (1A8), CD45.1 (A20), CD45.2 (104), MHC class II (I-A/I-E) (M5/114.15.2), SiglecF (E50-2440) (BD Biosciences), CD14 (Sa14-2) (BioLegend), CD68 (FA-11) (AbD Serotec), and MerTK (108928) (R&D Systems), as previously described1 (link), evaluated by flow cytometry using a FACSCaliber or FACSCanto flow cytometers (both from BD Biosciences, San Jose, CA), and the results were analyzed using CellQuest, FACSDiva (BD Biosciences), or FlowJo software (Tree Star). For intracellular staining of CD68, Leucoperm (AbDSerotec) was used as directed by the manufacturer.
Suzuki T., Arumugam P., Sakagami T., Lachmann N., Chalk C., Sallese A., Abe S., Trapnell C., Carey B., Moritz T., Malik P., Lutzko C., Wood R.E, & Trapnell B.C. (2014). Pulmonary Macrophage Transplantation Therapy. Nature, 514(7523), 450-454.
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2

Tissue Sectioning and Immunofluorescence Staining

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Tissue was embedded in optimal cutting temperature medium (TissueTec) and directly snap frozen in isopentane cooled by dry ice. The tissue was then cut into 8- or 14-µm sections by a cryostat and mounted on superfrost glass slides, which were dried for at least 1 h before storage at −70°C. Tissue sections were fixed and permeabilized with ice-cold acetone. The following antibodies were used: B220 (RA3-6B2; BioLegend), CD3 (pc; Abcam), CD68 (FA-11; AbD Serotec), CD103 (2E7; BioLegend), CD138 (281-2; BioLegend), F4/80 (BM8; AbD Serotec), IgG (STAR120F; AbD Serotec), Ly-6G (1A8; BioLegend), MARCO (ED31; AbD Serotec), MOMA1 (Abcam), and PNA (Vector). Tissue was mounted with Prolong Gold mountant (Invitrogen), and images were captured using a confocal scanning microscope (DMI6000; Leica Biosystems) connected to a True Confocal scanner (SP5; Leica Biosystems).
Parsa R., Lund H., Georgoudaki A.M., Zhang X.M., Ortlieb Guerreiro-Cacais A., Grommisch D., Warnecke A., Croxford A.L., Jagodic M., Becher B., Karlsson M.C, & Harris R.A. (2016). BAFF-secreting neutrophils drive plasma cell responses during emergency granulopoiesis. The Journal of Experimental Medicine, 213(8), 1537-1553.
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3

Histological Analysis of Hepatic Inflammation

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Liver specimens (4μm), stained with hematoxylin and eosin (H&E), were analyzed blindly [21 (link), 22 (link)]. Primary mAb against mouse neutrophils (Ly-6G) (1A8; BD Biosciences, San Jose, CA) and macrophages (CD68) (FA-11; AbD Serotec, Raleigh, NC) were used. Labeled cells were counted in 10 high-power fields (HPF). Superoxide levels in hepatic tissue were detected with ROS-sensing dye dihydroethidium (DHE; AnaSpec, Fremont, CA).
Zhang C., Zhang Y., Liu Y., Liu Y., Kageyama S., Shen X.D., Gao F., Zheng S., Busuttil R.W., Shaw G.D., Ji H, & Kupiec-Weglinski J.W. (2017). A SOLUBLE FORM OF PSGL-1 REQUIRES NRF2 SIGNALING TO PROTECT LIVER TRANSPLANT ENDOTHELIAL CELLS AGAINST ISCHEMIA-REPERFUSION INJURY. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 17(6), 1462-1475.
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4

Multiparametric analysis of myeloid cell populations

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BMDM were blocked with anti-CD16/32 (2.4G2; UCSF Monoclonal Antibody Core), stained with antibodies to myeloid surface markers, and analyzed on an LSRII (BD Biosciences). Intracellular staining was performed after 4 h incubation (for BMDM) or 2 h incubation (for peritoneal macrophages) with GolgiPlug (BD) and monensin (eBioscience), using Cytofix/Cytoperm (BD) according to manufacturer protocols. Results were analyzed using FACSDiva (BD) and FlowJo (TreeStar) software. Antibodies to the following were used: Ly6C (clone HK1.4), CD11c (N418), IL-1β (NJTEN3), TNF (TN3-19), rat IgG1 isotype (eBRG1), all eBioscience; Ly6G (1A8), Biolegend; Ly6B.2 (7/4), Cedarlane; CD11b (M1/70), F4/80 (BM8), Gr-1 (RB6-8C5), all UCSF Monoclonal Antibody Core; and CD68 (FA-11), ABD Serotec. Myeloid populations were delineated as previously described (16 (link)): granulocytes (CD11b+ Ly6G+), eosinophils (CD11b+ Ly6cint SSChi), monocytes (CD11b+ F4/80int SSClo Ly6c as indicated), red pulp macrophages (F4/80hi CD11b−), conventional DC (CD11chi), and plasmacytoid DC (CD11cint Siglec H+).
Fontana M.F., Baccarella A., Pancholi N., Pufall M.A., Herbert D.R, & Kim C.C. (2014). JUNB is a Key Transcriptional Modulator of Macrophage Activation. Journal of immunology (Baltimore, Md. : 1950), 194(1), 177-186.
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5

Liver Macrophage Quantification Protocol

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Liver specimens (4μm), stained with hematoxylin/eosin (H&E) were analyzed blindly by modified Suzuki's criteria (16 (link)). Primary mAb against mouse macrophages CD68 (FA-11; AbD Serotec, Raleigh, NC) was used. Liver sections were evaluated blindly by counting labeled cells/10 HPF.
Ji H., Liu Y., Zhang Y., Shen X.D., Gao F., Busuttil R.W., Kuchroo V.K, & Kupiec-Weglinski J.W. (2014). T-Cell Immunoglobulin and Mucin Domain 4 (TIM-4) Signaling in Innate Immune-Mediated Liver Ischemia-Reperfusion Injury. Hepatology (Baltimore, Md.), 60(6), 2052-2064.
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6

Histopathological Assessment of Liver Injury

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Perfused livers were fixed with 4% paraformaldehyde for 24 hours and then embedded in paraffin. For detection of liver necrosis and apoptosis, formalin-fixed liver tissue sections were stained with hematoxylin and eosin (H&E) for evaluation of necrosis. For apoptosis detection, tissue sections were stained with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP digoxigenin nick-end labeling (TUNEL) according to the manufacturer’s instructions (In situ cell death detection kit, POD, Roche). For immunohistochemical detection of infiltrating inflammatory cells, primary mAbs against mouse Ly-6G (1A8; BD Biosciences, San Jose, CA) and CD68 (FA-11; AbD Serotec, Raleigh, NC) were used on liver specimens, and positive cells were counted in 15 high power fields (HPF)/section for quantification of infiltrating immune cells.
Liu X., Hu Z., Zhou B., Li X, & Tao R. (2015). Chinese Herbal Preparation Xuebijing Potently Inhibits Inflammasome Activation in Hepatocytes and Ameliorates Mouse Liver Ischemia-Reperfusion Injury. PLoS ONE, 10(7), e0131436.
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7

Liver Histology and Immunohistochemistry Protocol

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Liver histology and immunohistochemistry were performed as previously published.[14 (link), 15 (link)] Liver specimens embedded in paraffin were processed for hematoxylin and eosin (H&E) staining. Liver steatosis was assessed by Oil Red O staining. Immunohistochemistry was performed using CD3 (HIT3a), Ly-6G (1A8), and PECAM-1 (MEC13.3) from BD Biosciences, CD68 (FA-11, Serotec), MMP-9 (AF909; R&D Systems), vWF (A0082, DAKO), PCNA (PC10; Neomarkers) and pH3 (Ser10; Cell Signaling) antibodies at optimal dilutions. Dual/triple staining was detected by immunofluorescence with Alexa Fluor 594 (red) and Alexa Fluor 488 (green) labeled secondary antibodies (Molecular Probes). Alexa Fluor 488 phalloidin and Vectashield mounting media with DAPI (Vector Laboratories) were used for F-actin and nuclear staining, respectively. Sections were blindly evaluated by counting 10 high-powered fields (HPFs)/section in triplicate. The proliferation index is expressed as the percentage of PCNA, or pH3 stained hepatocytes per total number of hepatocytes.
Kato H., Kuriyama N., Duarte S., Clavien P.A., Busuttil R.W, & Coito A.J. (2014). MMP-9 DEFICIENCY SHELTERS ENDOTHELIAL PECAM-1 EXPRESSION AND ENHANCES REGENERATION OF STEATOTIC LIVERS AFTER ISCHEMIA AND REPERFUSION INJURY. Journal of hepatology, 60(5), 1032-1039.
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8

Purification and Immunophenotyping of BAL Cells

Cited 2 times
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BAL cells were purified by centrifugation on Percoll to remove surfactant and debris 31 (link). BAL cells or BMDMs were immunostained to detect CD115 (AFS98), F4/80 (BM8) (eBioscience), CD3 (145-2C11), CD11b (M1/70), CD11c HL3), CD16/32 (2.4G2), CD19 (1D3), CD64 (X54-5/7.1.1), CD131 (JORO50), Ly6G (1A8), CD45.1 (A20), CD45.2 (104), MHC class II (I-A/I-E) (M5/114.15.2), SiglecF (E50-2440) (BD Biosciences), CD14 (Sa14-2) (BioLegend), CD68 (FA-11) (AbD Serotec), and MerTK (108928) (R&D Systems), as previously described1 (link), evaluated by flow cytometry using a FACSCaliber or FACSCanto flow cytometers (both from BD Biosciences, San Jose, CA), and the results were analyzed using CellQuest, FACSDiva (BD Biosciences), or FlowJo software (Tree Star). For intracellular staining of CD68, Leucoperm (AbDSerotec) was used as directed by the manufacturer.
Suzuki T., Arumugam P., Sakagami T., Lachmann N., Chalk C., Sallese A., Abe S., Trapnell C., Carey B., Moritz T., Malik P., Lutzko C., Wood R.E, & Trapnell B.C. (2014). Pulmonary Macrophage Transplantation Therapy. Nature, 514(7523), 450-454.
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9

Immunohistochemistry of Amyloid Beta in Hippocampus

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Brains were cut into 10 micron sections using a cryostat. A series of sections through the hippocampus was taken, and the site of injection was located by staining for CD11B; this allowed identification of the antibody-injected brain region to use for further immunohistochemistry. Immunohistochemistry was performed as described before [27 (link)] using CD11B (5C6, 1:500, Serotec), CD68 (FA11, 1:500, Serotec) and Aβ (3D6, 1:1000, in house), FITC sheep anti-mouse F(ab)2 (1:500, Sigma), horse anti-mouse biotinylated (1:250, Vector) and rabbit anti rat biotinylated (1:250, Vector).
Fuller J.P., Stavenhagen J.B., Christensen S., Kartberg F., Glennie M.J, & Teeling J.L. (2015). Comparing the efficacy and neuroinflammatory potential of three anti-abeta antibodies. Acta Neuropathologica, 130(5), 699-711.
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10

Immunohistochemical Analysis of Liver Macrophages

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Liver specimens were embedded in Tissue Tec OCT compound (Miles, Elkhart, IN) and snap frozen in liquid nitrogen and immunostaining performed on cryostat sections (0.5 µm) as described previously [19] (link). Appropriate rat primary antibodies against mouse macrophage antigen-1 (Mac-1; M1/70), CD68 (FA-11, Serotec), Ly-6G (1A8) (BD Biosciences, San Diego, CA), MMP-9 (AF909; R&D Systems, Minneapolis, MN) were used at optimal dilutions. Primary antibody was replaced with dilution buffer for negative controls. Bound primary antibodies were detected using biotinylated anti-rat IgG and streptavidin peroxidase–conjugated complexes (Vector Laboratories, Burlingame, CA). Sections from inflammatory tissues known to be positive for each stain were included as positive controls. Sections were evaluated blindly by counting 30 high-power fields per section.
Duarte S., Kato H., Kuriyama N., Suko K., Ishikawa T.O., Busuttil R.W., Herschman H.R, & Coito A.J. (2014). Hepatic Ischemia and Reperfusion Injury in the Absence of Myeloid Cell-Derived COX-2 in Mice. PLoS ONE, 9(5), e96913.
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