Fa 11

Manufactured by Bio-Rad

Sourced in United Kingdom

The FA-11 is a compact benchtop centrifuge designed for general-purpose applications in the laboratory. It features a fixed-angle rotor that can accommodate various sample tube sizes. The centrifuge provides consistent and reliable performance for a wide range of sample preparation and separation tasks.

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Lab products found in correlation

M1 70, by BD (2 mentions) Fc blocker 2.4 g2, by BD (1 mentions) Cytomics fc500, by Beckman Coulter (1 mentions) Streptavidin pe, by Thermo Fisher Scientific (1 mentions) Bz x710 fluorescence microscope, by Keyence (1 mentions) Bz x analyzer software, by Keyence (1 mentions) Tissue tek, by Sakura Finetek (1 mentions) Z033429 2, by Abcam (1 mentions) Pg m1, by Agilent Technologies (1 mentions) Af953, by R&D Systems (1 mentions) Image pro plus 7.0, by Adobe (1 mentions) Immpact dab peroxidase substrate, by Vector Laboratories (1 mentions) Laminin, by Abcam (1 mentions) Vectastain abc hrp kit, by Vector Laboratories (1 mentions) Anti ly6g fitc, by BD (1 mentions) Zen software, by Zeiss (1 mentions) Af965, by R&D Systems (1 mentions) Af1029, by R&D Systems (1 mentions) Mabg mdect, by InvivoGen (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

CD68 (FA-11; (14 citations) Clone FA-11 (3 citations) Anti-CD68 (clone FA-11, (3 citations) Monoclonal rat anti-CD68 antibody (Clone FA-11) (2 citations) Anti-CD68 mAb (clone FA-11, (2 citations) CD68 (clone FA-11) (2 citations) Biotin-conjugated anti-CD68 Ab (FA-11, (2 citations)

10 protocols using fa 11

Kidney Leukocyte and Apoptosis Analysis

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Immunoperoxidase staining for leukocytes was performed on 2% paraformaldehyde-lysine-periodate-fixed kidney cryostat sections (4μm) using anti-CD68 (FA-11, Serotec, Oxford, UK) and anti-CD3 (KT3, Abcam) antibodies according to established protocols.[5 (link)] To assist leukocyte counting, cell nuclei were weakly stained with hematoxylin. The number of macrophages (CD68+) and T-cells (CD3+) was assessed in 20 glomeruli per kidney section. Immunoperoxidase staining for apoptotic cells was performed on methylcarn-fixed paraffin-embedded kidney sections (4μm) using an antibody to activated caspase-3.[5 (link)] The number of apoptotic cells (activated caspase-3+) in cortical tubules was assessed in each animal and recorded as apoptotic cells per 1000 tubule cross-sections (tcs). All scoring was performed on blinded slides.

Ma F.Y., Han Y., Nikolic-Paterson D.J., Kolkhof P, & Tesch G.H. (2015). Suppression of Rapidly Progressive Mouse Glomerulonephritis with the Non-Steroidal Mineralocorticoid Receptor Antagonist BR-4628. PLoS ONE, 10(12), e0145666.

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Identification of Kupffer Cells and FasL Expression

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The MNCs were incubated for 15 min at 4 °C with Fc-blocker (2.4 G2; BD PharMingen, USA) to prevent any nonspecific binding. For identification of Kupffer cells, MNCs were stained with a FITC labeled anti-F4/80 antibody (Ab) (BM8, eBioscience, San Diego, CA. USA), PE-Cy5 labeled CD11b Ab (M1/70, BD PharMingen), and biotin labeled CD68 Ab (FA-11, SEROTEC, Oxford, UK) and developed with PE-streptavidin (eBioscience). For analysis of FasL expression, a PE labeled FasL antibody (Kay-10, eBioscience) was used. Flow cytometric analysis was performed using a Cytomics FC500 instrument (Beckman Coulter, Indianapolis, IN, USA).

Nakashima H., Nakashima M., Kinoshita M., Ikarashi M., Miyazaki H., Hanaka H., Imaki J, & Seki S. (2016). Activation and increase of radio-sensitive CD11b+ recruited Kupffer cells/macrophages in diet-induced steatohepatitis in FGF5 deficient mice. Scientific Reports, 6, 34466.

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Immunohistochemical Analysis of CD68+ Macrophages

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Mice were euthanized with CO₂ and hind paw skin was dissected and fixed for 4 hours in 4% PFA, followed by overnight cryoprotection in sucrose (30% in PBS, pH 7.4) at 4 °C. The tissues were embedded in optimal cutting temperature medium (Tissue-Tek®, Sakura Finetek, Torrance, CA) and were flash-frozen in cold isopentane. Three series of transversal sections (thickness 8 μm) were prepared using a cryostat. After rinsing with PBS, the sections were blocked and permeabilized with 0.1% Tween-20 in PBS containing 5% normal donkey serum for 30 min at room temperature. Immunostaining was performed by overnight incubation at 4 °C with an anti-CD68 primary antibody (Serotec, FA-11, 1:200, #MCA1957), followed by incubation for one hour at room temperature with a secondary anti-rat Alexa Fluor 594 donkey antibody (Invitrogen cat. #A21209; 1:200). Nuclei were stained with DAPI (1:250 in PBS) for 10 min at room temperature and slides were mounted with fluorescence-safe mounting medium. Images were obtained at 10x or 40x magnification using a Keyence BZ-X710 fluorescence microscope coupled with BZ-X Analyzer software (Keyence).

Fotio Y., Mabou Tagne A., Squire E., Lee H.L., Phillips C.M., Chang K., Ahmed F., Greenberg A.S., Villalta S.A., Scarfone V.M., Spadoni G., Mor M, & Piomelli D. (2024). NAAA-regulated lipid signaling in monocytes controls the induction of hyperalgesic priming in mice. Nature Communications, 15, 1705.

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Immunohistochemical Analysis of Muscle Fiber Composition

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Transverse sections of 8 μm were fixed in ice-cold acetone for 8 min. For CD11b (rat monoclonal M1/70, 1:300, BD Pharmingen), CD68 (rat monoclonal FA-11, 1:100, AbD Serotec), and laminin γ1 chain (rabbit polyclonal 1083 + E1, 1:100, kindly provided by Dr. T. Sasaki), sections were blocked in 3% BSA in PBS at RT for 30 min followed by incubation in primary antibody at RT for 1 h. For embryonic myosin heavy chain (mouse monoclonal F1.652, 1:10, Developmental Studies Hybridoma Bank), sections were blocked in 4% goat serum and 0.05% Triton-X in PBS at RT for 40 min followed by incubation in primary antibody at RT for 90 min. Primary antibodies were incubated with appropriate secondary antibodies for 60 or 45 min (embryonic myosin heavy chain). Antibody stained sections were analyzed using a Zeiss Axioplan fluorescent microscope using Openlab 3 and an ORCA 1394 ER digital camera. The percentage of embryonic myosin heavy chain–positive fibers was obtained by counting the number of fibers positive for embryonic myosin heavy chain in a whole quadriceps section and dividing by the total number of myofibers.

Holmberg J., Alajbegovic A., Gawlik K.I., Elowsson L, & Durbeej M. (2014). Laminin α2 Chain-Deficiency is Associated with microRNA Deregulation in Skeletal Muscle and Plasma. Frontiers in Aging Neuroscience, 6, 155.

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Immunofluorescence and IHC Staining Protocols

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The following primary antibodies were used for immunofluorescence staining at the indicated dilutions: anti-GFAP (1:1000, Dako, Z033429-2), anti-F4/80 (1:50, Abcam, AB6640), human anti-Cathepsin B (1:1000, R&D, AF953), murine anti-Cathepsin B (1:500, R&D, AF965), human anti-CD68 (1:1, Dako, PG-M1), and murine anti-CD68 (1:3000, Serotec, FA-11). The following primary antibodies were used for IHC at the indicated dilutions: human anti-CD163 (1:200, Vector, 10D6), human anti-CD68 (1:200, Dako, M0876), anti-collagen IV (1:200, Vector labs, PK6101), murine anti-GFP (1:100, Abcam, AB1218), and human anti-CCR2 (1:400, Novus biologicals, E68). JPM-OEt (100 mg/kg/day) was administered intraperitoneally in 30% DMSO/70% PBS as previously described (19 (link)).

Bakst R.L., Xiong H., Chen C.H., Deborde S., Lyubchik A., Zhou Y., He S., McNamara W., Lee S.Y., Olson O.C., Leiner I.M., Marcadis A.R., Keith J.W., Al-Ahmadie H.A., Katabi N., Gil Z., Vakiani E., Joyce J.A., Pamer E, & Wong R.J. (2017). Inflammatory monocytes promote perineural invasion via CCL2-mediated recruitment and cathepsin B expression. Cancer research, 77(22), 6400-6414.

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Immunohistochemical Staining and Quantification

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Using VECTASTAIN ABC-HRP Kits (Vector Labs), 10 µm sections were stained with antibodies against CD68 (biotinylated clone FA-11, 1:10, AbD Serotec), CD31 (1:100, BD Pharmingen), laminin (1:300, Abcam), α-actin (clone ab5694 1:200, Abcam), or anti-TSP-1 Ab4 (clone 6.1 1:100, Thermo). Secondary antibodies were included in the species-specific kit, followed by ImmPACT DAB peroxidase substrate (Vector Labs). Slides were scanned using Leica SCN400 or Aperio AT2 at 20X magnification. Positive staining in the images was quantified using Photoshop CS2 (Adobe) and Image Pro Plus (7.0).

Gajeton J., Krukovets I., Muppala S., Verbovetskiy D., Zhang J., & Stenina‐Adognravi O. (2020). Hyperglycemia-Induced MIR-467 Drives Tumor Inflammation And Growth In Breast Cancer.

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Liver Assessment and Cell Quantification

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Hematoxylin/eosin stained liver sections were assessed by Suzuki’s score. Liver specimens, stained with Alexa Fluor 488 anti-CD68 (FA-11; 5μg/ml, BIO-RAD, Hercules, CA), FITC anti-Ly-6G (1A8; 5μg/ml, BD Biosciences, San Jose, CA), and PE anti-CDK2 (78B2; 5μg/ml, Cell Signaling Technology, Danvers, MA) were analyzed blindly.

Xu J., Xue Z., Zhang C., Liu Y., Busuttil R.W., Zhang J., Kupiec-Weglinski J.W, & Ji H. (2019). Inhibition of Cyclin-dependent Kinase 2 Signaling Prevents Liver Ischemia and Reperfusion Injury. Transplantation, 103(4), 724-732.

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Immunofluorescence Labeling of Heart Valve Tissue

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For immunofluorescence labeling, 5 µm thick cryosections from heart valve tissue of all experimental groups were fixed in ice-cold acetone for 10 min followed by ice-cold 80 % methanol for 5 min. The rat-anti-mouse monoclonal antibodies of 30-F11 (BD Biosciences, Heidelberg, Germany) and FA-11 (Bio-Rad Laboratories GmbH, Muenchen, Germany) were applied for CD45 and F4/80 detection respectively, and allowed to incubate for 60 min at room temperature. The cryosections incubated with the rat-anti-mouse monoclonal antibodies were rinsed three times in TBS-T washing buffer and were then incubated with Cy3-conjugated AffiniPure Donkey Anti-Rat IgG (Jackson Immunoresearch Laboratories Inc., Pennsylvania, USA) for 45 min at room temperature. After rinsing in TBS-T buffer and distilled water, sections were mounted in Vectashield H1200 mounting medium containing DAPI (Linaris biologische Produkte GmbH, Wertheim-Bettingen, Germany) and stored at −20 °C. Antibody specificity control staining was performed in accordance but by omitting the primary antibodies. Immunofluorescence labeling was analyzed with the LSM 9000 (Aiyscan 2) microscope using the ZEN software (both Carl Zeiss, Germany).

Schwarz C., Töre Y., Hoesker V., Ameling S., Grün K., Völker U., Schulze P.C., Franz M., Faber C., Schaumburg F., Niemann S, & Hoerr V. (2021). Host-pathogen interactions of clinical S. aureus isolates to induce infective endocarditis. Virulence, 12(1), 2073-2087.

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Immunohistochemistry of Microglia and Astrocytes

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30μm microtome-cut sagittal sections were incubated in X34 for one hour at room temperature in 60% PBS/40% EtOH mix; pH was adjusted with 1N NaOH. Following this, sections were washed briefly with 60% PBS/40% EtOH before blocking in 5% donkey or goat serum for 1h at room temperature. Primary antibodies were diluted in blocking buffer and incubated at 4°C overnight with slow agitation unless mentioned otherwise. Primary antibodies were used as follows: IBA1 (1:500; 019-19741, Fujifilm or NB100-1028, Novus Biologicals), CD68 (1:100; FA-11, BioRad), P2RY12 (1:100 at room temperature, S16007D, BioLegend), TMEM119 (1:500; E3E1O, Cell Signaling Technology), Clec7a (1:50 at room temperature; mabg-mdect, InvivoGen), GFAP (1:2000; 2E1.E9 Alexa Flour 488-conjugated, BioLegend), 6E10 (1:1000; Alexa Flour 647-conjugated, BioLegend), NeuN (1:2000; ab104224, Abcam), LAMP1/CD107a (1:500; AF4320, Novus Biologicals), CathepsinB (1:50; AF965, R&D Systems), Cathepsin D (1:50; AF1029, R&D Systems). The next day, sections were washed and incubated with secondary antibody diluted in blocking buffer, followed by a 4′,6-Diamidin-2-phenylindol (DAPI, 5μg/mL) where appropriate before mounting sections onto slides (Prolong^™ Gold Antifade reagent, Thermo Fisher Scientific).

Sharma M., Ellis R.L, & Hinton D.M. (1992). Identification of a family of bacteriophage T4 genes encoding proteins similar to those present in group I introns of fungi and phage.. Proceedings of the National Academy of Sciences of the United States of America, 89(14), 6658-6662.

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Macrophage Quantification in Muscle Cryosections

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Muscle cryosections (5 mm) were stained for the presence of macrophages, using anti-mouse anti-F4/80 antibody (Ab; BM-8; Invitrogen, Carlsbad, CA) and anti-mouse CD68 Ab (FA-11; Bio-Rad AbD Serotec, Raleigh, NC), as previously detailed. 18 To combine immunostaining with histopathologic analysis, we counterstained with hematoxylin and eosin. To determine the number of proliferating macrophages, we stained cryosections of muscle with anti-mouse CD68 Ab, and anti-mouse Ki-67 Ab (SP6; Vector Laboratories, Burlingame, CA), followed by fluorescein isothiocyanate (FITC)-conjugated goat anti-rat IgG Ab (Invitrogen) and cyanine (Cy)3-conjugated goat anti-rabbit IgG Ab (Invitrogen). We enumerated the number of CD68 þ /Ki-67 þ cells per field in the entire cross section.

Baek J.H., Many G.M., Evesson F.J, & Kelley V.R. (2017). Dysferlinopathy Promotes an Intramuscle Expansion of Macrophages with a Cyto-Destructive Phenotype. The American journal of pathology, 187(6).

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