Anti cd68

Manufactured by Merck Group

Sourced in United States

Anti-CD68 is a laboratory reagent used for the detection and characterization of cells that express the CD68 antigen, which is primarily found on macrophages and monocytes. It is a useful tool for immunohistochemistry and flow cytometry applications.

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Lab products found in correlation

Ab5076, by Abcam (1 mentions) Duo82040, by Merck Group (1 mentions) Anti iba1, by Abcam (1 mentions) Alexa fluor 555, by Thermo Fisher Scientific (1 mentions) Anti cd45, by Agilent Technologies (1 mentions) Target retrieval solution ph 9, by Agilent Technologies (1 mentions) Ultra cc1, by Roche (1 mentions) Anti cd20, by Agilent Technologies (1 mentions) Anti pan cytokeratin, by Agilent Technologies (1 mentions) Anti α sma, by Abcam (1 mentions) 3 3 diaminobenzidine (dab), by Merck Group (1 mentions) Anti βiii tubulin, by Merck Group (1 mentions) Facsaria 3 cytometer, by BD (1 mentions) Anti nestin, by Santa Cruz Biotechnology (1 mentions) Anti gfap, by Agilent Technologies (1 mentions) Trypsin, by Merck Group (1 mentions) Biotinylated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Streptavidin hrp, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Rabbit anti-CD68 (2 citations) Anti-CD68 antibody (2 citations) Anti-CD68 ED-1 mouse monoclonal (2 citations)

7 protocols using anti cd68

Immunofluorescence Staining of Microglia

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The cells were treated with 4% paraformaldehyde for 10 min, 0.1% TritonTM X-100 for 15 min, and blocked with 1%BSA for 1 h at room temperature. The cells were then incubated with the primary antibodies (anti-IBA1 (1:200; AB5076, Abcam, Cambridge, UK) and anti-CD68 (1:100; 14-0681-82, Sigma-Aldrich, St. Louis, MO, USA)) for an overnight period at 4 °C. The cells were then washed with PBS-T and incubated with the secondary antibodies (Alexa Fluor@555 anti-rat IgG (1:2000; 4417s, CST, Danvers, MA, USA), Alexa Fluor 488 (A-11055, 1:2000; Thermo Fisher Scientific, Thermo Fisher Scientific, Waltham, MA, USA), and Alexa Fluor 555 (A32816, 1:2000; Thermo Fisher Scientific, Thermo Fisher Scientific, Waltham, MA, USA)) at room temperature for 1 h.
After being washed three times for 10 min with PBS-T, DAPI (1:10,000; DUO82040, Sigma-Aldrich, St. Louis, MO, USA) was used as a counterstain on the cells. Finally, fluorescence images were captured with a fluorescence microscope (Nikon, A1, Shanghai, China), and the analysis of the fluorescence images was performed by ImageJ2.

Li C., Wu Y., Huang M.Y, & Song X.J. (2023). Characterization of Inflammatory Signals in BV-2 Microglia in Response to Wnt3a. Biomedicines, 11(4), 1121.

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Immunofluorescence Staining of Brain Cells

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For immunofluorescence staining, brains of each group were sectioned into 20-mm slices and permeabilized with 0.5% Triton X-100 for 1 hour at room temperature, then blocked for 1 hour using 1% bovine serum albumin and 10% goat serum in phosphate-buffered saline. Afterward, sections were incubated in the following antibodies: anti-CC1 (1:500, Cell Signaling Technology, Danvers, Mass) for oligodendrocytes (OLs), anti-CD68 (Sigma, St Louis, Mo) for microglia, anti-ionized calcium-binding adapter molecule 1 (1:500, Sigma) for M1, anti-arginase 1 (Arg1; 1:500, Sigma) for M2, and anti-myelin basic protein (MBP; 1:400, Biolegend, San Diego, Calif); then, sections were diluted in blocking buffer at 4 C overnight. Next, sections were washed 3 times with phosphate-buffered saline, then incubated with Alexa Fluor 488-conjugated goat anti-mouse immunoglobulin (Ig)G/IgM (1:400; Invitrogen, Carlsbad, Calif), Alexa Fluor

Liu G., Yan Y., Shi B., Huang J., Mu H., Li C., Chen H, & Zhu Z. (2020). Benefits of progesterone on brain immaturity and white matter injury induced by chronic hypoxia in neonatal rats. The Journal of thoracic and cardiovascular surgery, 160(2).

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Immunohistochemical Characterization of iehAM

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Explanted iehAM were fixed in 4% formaldehyde, embedded in paraffin, and stained with hematoxylin and eosin according to standard protocols. Additional sections were deparaffinized and pretreated with Ultra CC1 (Ventana Medical Systems, Tucson, AZ, USA) for antigen retrieval for anti-CD45, anti-CD20, anti-CD68, and anti-CD3 antibody staining and with target retrieval solution pH 9 (Dako, Heverlee, Belgium) for anti-pan-cytokeratin and anti-GFAP antibody staining. After endogenous peroxidase blocking, sections were incubated with 1:25 anti-CD45 (Dako, Heverlee, Belgium, clone 2B11 + PD7/26), 1:200 anti-CD20 (Dako, clone L-26), 1:1000 anti-CD68 (Merck, Darmstadt, Germany), 1:100 anti-pan-cytokeratin (Dako, clone 6F2), 1:200 anit-GFAP (Dako, clone AE1/AE3), and 1:20 anti-CD3 (Monosan, Uden, Netherlands, clone PS-1) antibodies. After incubation with peroxidase-labelled secondary antibodies, sections were visualized with a chromogen DAB (3,3′-diaminobenzidine) solution.

Hillenmayer A., Wertheimer C.M., Gerhard M.J., Priglinger S.G., Ohlmann A, & Wolf A. (2023). Epiretinal Amniotic Membrane in Complicated Retinal Detachment: a Clinical and In Vitro Safety Assessment. Ophthalmology and Therapy, 12(3), 1635-1648.

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Immunohistochemical Analysis of Aortic Tissue

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Fixed and paraffin-embedded aorta tissue samples were cut into 4-µm sections and were dewaxed. Endogenous peroxidase activity and non-specific binding were blocked by incubation with 3% hydrogen peroxide and 100% non-immune serum (closed serum; cat. no. G9023; Sigma-Aldrich; Merck KGaA) respectively for 30 min at room temperature. Sections were incubated with anti-CD68 (EMD Millipore; cat. no. 051050; 1:250) and anti-α-SMA (Abcam; cat. no. ab32575; 1:250) antibodies at 4°C overnight, and with anti-rabbit secondary antibody (1:5,000; cat. no. ab181658; Abcam) at room temperature for 1 h. Diaminobenzidine hydrochloride (Dako; Agilent Technologies, Inc.) was then added to localize positive staining that was acquired using light microscopy. The sections were counterstained with hematoxylin for 10 min at room temperature. The data were analyzed via densitometry using ImageJ software (version 146; National Institutes of Health).

Zhao Y., Li W, & Zhang D. (2021). Gycyrrhizic acid alleviates atherosclerotic lesions in rats with diabetes mellitus. Molecular Medicine Reports, 24(5), 755.

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Immunohistochemical Analysis of Renal Tissue

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Histological sections of renal tissue were incubated for 1 h at room temperature with the following antibodies: anti-V1a (1:200; CUSABIO, College Park, MD, USA), anti-CD68 (1:200, Millipore, Billerica, MA, USA), anti-angiotensin (ANG) II type 1 (anti-AT-1, 1:50; Research Diagnostics Inc, Flanders, NJ, USA), anti-CD43 (1:50; Seralab, Crawley Down, UK), anti-Toll-like receptor 4 (anti-TLR4, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA), and anti-uromodulin (1:100; Millipore, Billerica, MA, USA). The reaction product was detected with horseradish peroxidase-conjugated HRP System (Anti-rabbit Polymer; Dako, Glostrup, Denmark), and the color reaction was developed with 3,3-diaminobenzidine (Sigma-Aldrich, St. Louis, MO, USA). The histological sections were divided into 20 fields (0.087 mm² each; magnification, × 400), and the mean value for each animal was presented and calculated with GraphPad Prism, version 5.03 (GraphPad Software Inc., San Diego, CA, USA).

Castro L.U., Otsuki D.A., Sanches T.R., Souza F.L., Santinho M.A., da Silva C., Noronha I.D., Duarte-Neto A.N., Gomes S.A., Malbouisson L.M, & Andrade L. (2022). Terlipressin combined with conservative fluid management attenuates hemorrhagic shock-induced acute kidney injury in rats. Scientific Reports, 12, 20443.

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Spinal Cord Cell Isolation and Flow Cytometry

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Six weeks after the implantation, the animals’ spinal cords were isolated. A 1-cm fragment was collected, including the epicenter of the lesion and equal rostral end caudal portions. The non-degraded PLGA membranes were removed and the tissue was mechanically and enzymatically dissociated with trypsin (Sigma-Aldrich). The cell suspension was fixed for 30 min with 4% paraformaldehyde and subsequently blocked for 30 min with 3% BSA in PBST. After blocking for 20 min with 3% BSA in PBS with 0.1% Triton X-100, the cells were incubated with primary antibodies, including anti-GFAP (DAKO; 14.5 μg/mL), anti-βIII tubulin (Millipore, 05559), anti-nestin (Santa Cruz, SC-33677, 1 μg/mL), and anti-CD68 (Millipore, MAB 1435). The cells were washed twice with PBS1X and incubated for 1 h with the secondary antibody Alexa-fluor 488 anti-mouse or anti-rabbit (10 μg/mL, Thermo Fisher Scientific, USA) at 37°C. Negative controls (samples incubated only with the secondary antibody) were included for setting up the machine voltages and to determine the negative population. The cells were analyzed using a FACSAria III cytometer (Becton Dickinson Biosciences, USA), equipped with a 488 nm argon laser and the FACSDiva 6.0 software. An average of 5×10⁴ events was analyzed.

Reis K.P., Sperling L.E., Teixeira C., Sommer L., Colombo M., Koester L.S, & Pranke P. (2020). VPA/PLGA microfibers produced by coaxial electrospinning for the treatment of central nervous system injury. Brazilian Journal of Medical and Biological Research, 53(4), e8993.

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Immunohistochemical Analysis of Liver Macrophages

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Liver tissue was fixed in 10% neutral buffered formalin and paraffin embedded, and cut into 4 μm sections. Sections were stained with H&E. For immunohistochemical analysis, paraffin-embedded sections were used for immunostaining for cell surface marker, CD68 (expressed particularly on monocytes/macrophages), as described previously^{45 (link), 46 (link)} with slight modification. Sections were deparaffinized and rehydrated, followed by incubation in 3% H₂O₂ for 10 min. After blocking with 5% BSA, the sections were incubated with anti-CD68 (1 : 100, Millipore, Bedford, MA, USA) overnight at 4 °C, followed by biotinylated secondary antibodies (1 : 200, Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 30 min and incubation with HRP-streptavidin (1 : 400, Invitrogen, Carlsbad, CA, USA) for 30 min. Color development was performed with DAB for 3–5 min.

Zhan Y., Wang Z., Yang P., Wang T., Xia L., Zhou M., Wang Y., Wang S., Hua Z, & Zhang J. (2014). Adenosine 5′-monophosphate ameliorates D-galactosamine/lipopolysaccharide-induced liver injury through an adenosine receptor-independent mechanism in mice. Cell Death & Disease, 5(1), e985-.

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