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Mca341r

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The MCA341R is a laboratory equipment product from Bio-Rad. It is a multi-channel analyzer designed for automated analysis of samples. The core function of the MCA341R is to perform simultaneous measurements and data processing for multiple samples.

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Lab products found in correlation

Mca342r, by Bio-Rad (3 mentions) Nis elements ar 4, by Nikon (2 mentions) Dxm1200c, by Nikon (2 mentions) Af942, by R&D Systems (2 mentions) Imagepro 6.0 plus, by Media Cybernetics (2 mentions) A21202, by Thermo Fisher Scientific (2 mentions) Ab6326, by Abcam (1 mentions) Mitomycin, by Merck Group (1 mentions) Ab5320, by Merck Group (1 mentions) N6 2 o dibutyryladenosine 3 5 cyclic monophosphate sodium salt dbcamp, by Merck Group (1 mentions) Z0311, by Agilent Technologies (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Leica tcs nt laser microscope, by Leica Microsystems (1 mentions) Ab5566, by Merck Group (1 mentions) Primo star microscope, by Zeiss (1 mentions) Sc 32757, by Santa Cruz Biotechnology (1 mentions) Sc 7271, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 555 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Normal mouse igg, by Agilent Technologies (1 mentions)

37 protocols using mca341r

1

Quantification of Glomerular Pathology

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Kidney samples fixed in formalin or methanol-Carnoy fixative solution were embedded in paraffin. Sections of 2–3 µm were stained with periodic acid-Schiff (PAS) reagent to assess glomerular hypercellularity, necrotizing lesions, and formation of glomerular crescents (crescentic glomeruli per 100 glomeruli was calculated and expressed as a percentage). Infiltrating leukocytes were immunohistochemically stained for ED1+ (catalog MCA341R AbD Serotec), CD169+, clone ED3, a marker for MI macrophages (catalog MCA343GA, AbD Serotec), and CD163+, clone ED2, a marker for M2 macrophages (catalog MCA342R, AbD Serotec) infiltrates. Slides were reacted with mouse anti-rat ED1, anti-rat CD169, and anti-rat CD163 and peroxidase-coupled anti-mouse IgG second antibodies (catalog P044701-2, Agilent/Dako) as described [6 (link),24 (link),35 (link)]. Positively stained cells per 100 glomeruli were counted and expressed per glomerular section. All quantitative morphological analyses were performed in a blinded fashion.
Garcia G.E., Lu Y.J., Truong L.D., Roncal-Jiménez C.A., Miyazaki M., Miyazaki-Anzai S., Cara-Fuentes G., Andres-Hernando A., Lanaspa M., Johnson R.J, & Leamon C.P. (2021). A Novel Treatment for Glomerular Disease: Targeting the Activated Macrophage Folate Receptor with a Trojan Horse Therapy in Rats. Cells, 10(8), 2113.
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2

Immunochemical Identification of CNS Cell Types

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Basic reagents were purchased from Sigma (St Louis, MO), unless indicated otherwise. Bromodeoxyuridine (BrdU; 5-bromo-2′-deoxyuridine) and TIMP-1 from human neutrophil granulocytes were purchased from Calbiochem (San Diego, CA); mitomycin and N6,2′-O-dibutyryladenosine-3′,5′-cyclic monophosphate sodium salt (dbcAMP) were purchased from Sigma; cholera toxin B (CTB) subunit was purchased from List Biological Laboratories (Campbell, CA); and 4′,6-diamidino-2-phenylindole (DAPI) was purchased from Molecular Probes (Eugene, OR). The antibodies used for immunodetection were as follows: monoclonal mouse anti-BrdU (B2531; Sigma) and polyclonal rabbit anti–glial fibrillary acidic protein (GFAP; Z0334; Dako, Carpinteria, CA; for dual labeling with NG2, Iba1, and GFAP); monoclonal rat anti-BrdU (ab6326; Abcam, Cambridge, MA; for dual labeling with O1); polyclonal goat anti-CTB (703; List Biological Laboratories); polyclonal rabbit anti-NG2 (AB5320; EMD Millipore, Billerica, MA); monoclonal mouse anti-O1 (MAB344; Chemicon, Temecula, CA); monoclonal mouse anti-CD11b (MCA618R; Serotec, Raleigh, NC); polyclonal rabbit anti-Iba1 (019-19741; Wako, Richmond, VA); monoclonal mouse rat anti-CD68 (MCA341R; Serotec); polyclonal goat anti–calcitonin gene–related peptide (CGRP; 1720-9007; Biogenesis, Bournemouth, United Kingdom); and polyclonal rabbit anti-S100 (Z0311; Dako).
Liu H., Angert M., Nishihara T., Shubayev I., Dolkas J, & Shubayev V.I. (2015). Spinal Glia Division Contributes to Conditioning Lesion–Induced Axon Regeneration Into the Injured Spinal Cord: Potential Role of Cyclic AMP–Induced Tissue Inhibitor of Metalloproteinase-1. Journal of neuropathology and experimental neurology, 74(6), 500-511.
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3

Immunocytochemistry of Bone Marrow-Derived Macrophages

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Matured bone marrow-derived macrophages from both strains were seeded at a density of 1 × 106 cells on a coverslip. Cells were then fixed in 3.7% paraformaldehyde for 10 minutes, washed with PBS, permeabilized with acetone for 5 minutes on ice, and blocked with goat serum for one hour at room temperature. Cells were washed twice with PBS and incubated with monoclonal mouse anti-rat CD68 (ED1) (MCA341R; Serotec, Madrid, Spain) for 1 hour at room temperature. After washing with PBS, slides were incubated with secondary fluorescent antibody (1:1000) (Alexa fluor 546 Goat Anti-mouse, highly cross absorbed, Molecular Probes, USA) for 30 minutes. The preparation was mounted with mowiol (Calbiochem, La Jolla, CA) and cells were viewed using a Leica TCS NT laser microscope (Leica Microsystems, Wetzlar, Germany), equipped with a PT APO objective (60 ×).
Jung M., Brüne B., Hotter G, & Sola A. (2016). Macrophage-derived Lipocalin-2 contributes to ischemic resistance mechanisms by protecting from renal injury. Scientific Reports, 6, 21950.
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4

Histological Analysis of Knee Joint

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Knee joints were fixed in 10% neutral buffered formalin for three days and then placed in a decalcification solution (ImmunoCal Cat # 1440, Decal Chemical Corp, Suffern, NY) for 5 days. On the fourth day, knees were trimmed along the sagittal axis approximately at the mid-trochlear level to separate medial and lateral condyles of the joint, and decalcification was pursued until completion. After sample dehydration and paraffin embedding, 5-µm-thick sections were cut on the lateral condyle and stained: hematoxylin and eosin (H&E) for the characterization of synovium and proteoglycan-containing cartilage identified by Safranin O/Fast green using a procedure adapted from Lillie and Fulmer19 and from Prophet et al20 . Macrophages and osteoclasts were detected with an anti-CD68 antibody (MCA341R, Serotec, Puchheim, Germany) applied on paraffin sections as described by Damoiseaux et al21 (link).
Accart N., Dawson J., Obrecht M., Lambert C., Flueckiger M., Kreider J., Hatakeyama S., Richards P.J, & Beckmann N. (2022). Degenerative joint disease induced by repeated intra-articular injections of monosodium urate crystals in rats as investigated by translational imaging. Scientific Reports, 12, 157.
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5

Quantifying Renal Macrophages and TRPV1

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The kidneys were fixed with cut into sections. The sections were blocked and incubated with rabbit anti-TRPV1 polyclonal antibody (1:500, AB5566, Millipore, Billerica, MA, USA) or mouse anti-CD68 (a macrophage marker) monoclonal antibody (1:100, MCA341R, AbD Serotec, Kidlington, UK) at 4 °C overnight, then incubated with Rhodamine Red (TM)-X donkey anti-rabbit IgG (1: 100, 711–295-152, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) or Rhodamine Red (TM)-X donkey anti-mouse IgG (1: 100, 715–295-152, Jackson ImmunoResearch Laboratories) for 1h at room temperature. Images were taken with a fluorescent microscope (Olympus BX41 model, Olympus Optical Co. Ltd, Tokyo, Japan). For the quantification of macrophage staining, 10 coronal sections through the renal hilum of each kidney at interval of 20 μm were selected for staining. For each section, 10 fields of view from cortex and medulla each were chosen randomly for counting the number of macrophages. The value was expressed as macrophage number per square millimeter.
Yu S.Q., Ma S, & Wang D.H. (2020). Ablation of TRPV1-positive nerves exacerbates salt-induced hypertension and tissue injury in rats after renal ischemia-reperfusion via infiltration of macrophages. Clinical and experimental hypertension (New York, N.Y. : 1993), 43(3), 254-262.
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6

Localization of Macrophage Markers in Kidney Tissue

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To study the specific localization of macrophage, ED-1, iNOS (inducible nitric
oxide synthase), and nitrotyrosine, sections of paraffin-embedded kidneys were
used. This procedure was performed as previously described19 (link)
-20 (link) and the method consisted of an
immunoperoxidase reaction. Tissue fragments were incubated overnight at 4°C with
a primary monoclonal anti-iNOS Ab (1:10, sc-7271, Santa Cruz Biotechnology, CA,
USA), or for 1 h at room temperature with a primary monoclonal anti-ED-1 Ab
(1:1000, MCA341R, Serotec, Oxford, UK) or primary monoclonal anti-nitrotyrosine
Ab (1:400, SC-32757, Santa Cruz Biotechnology, CA, USA). Quantitative analysis
of ED-1 macrophages and iNOS was performed in tissue sections from the
juxtamedullary segments in a blinded fashion, with counting being performed
using a Primo Star microscope (Carl Zeiss, Jena, Germany). For nitrotyrosine
analyses, one slide from each animal (n = 6 per group) was used, and 25 fields
in the juxtamedullary region of the kidney were observed to obtain an average
score, as previously described for histopathology.19 (link)
-20 (link)
Anselmo N.A., Paskakulis L.C., Garcias R.C., Botelho F.F., Toledo G.Q., Cury M.F., Carvalho N.Z., Mendes G.E., Iembo T., Bizotto T.S., Cury P.M., Chies A.B, & Carlos C.P. (2018). Prior intake of Brazil nuts attenuates renal injury induced by ischemia and reperfusion. Jornal Brasileiro de Nefrologia, 40(1), 10-17.
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7

Immunostaining of Mouse Macrophages

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ED-1 (mouse monoclonal antibody, MCA 341R, Serotec, Puchheim, Germany) and ED-2 (mouse monoclonal antibody, MCA 342R, Serotec). A rabbit polyclonal antibody against myeloperoxidase (MPO) was purchased from Dako (Hamburg, Germany). Fluorescent labelled antibodies Alexa Fluor 555 goat anti-rabbit or Alexa Fluor 488 goat anti-mouse were purchased from Invitrogen (Darmstadt, Germany).
Ahmad S., Ramadori G, & Moriconi F. (2018). Modulation of Chemokine- and Adhesion-Molecule Gene Expression and Recruitment of Neutrophil Granulocytes in Rat and Mouse Liver after a Single Gadolinium Chloride or Zymosan Treatment. International Journal of Molecular Sciences, 19(12), 3891.
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8

Immunohistochemical Analysis of Macrophage-like Synoviocytes

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The sections at each period were deparaffinized and soaked in 0.3% hydrogen peroxide. Cluster of differentiation 68 (CD68) was used as a marker of macrophage-like type A synoviocytes [24 (link)]. Endogenous peroxidase was inactivated with 3% H2O2 in PBS for 20 min at room temperature. The slides were incubated with mouse anti-rat CD68 antibody (MCA341 R, AbD Serotec, Raleigh, NC, USA; dilution, 1:400) for 24 h at 4 °C. Goat anti-mouse immunoglobulin (IgG) (Nichirei, Tokyo, Japan) for CD68 was used as the secondary antibody and incubated for 30 min at room temperature. The final detection step was carried out using 3,30-diaminobenzidine tetrahydrochloride (DAB) (Sigma, St. Louis, MO, USA) in 0.1 M imidazole and 0.03% H2O2 as the chromogen. Counterstaining was performed using Carazzi’s hematoxylin. For the negative control, normal mouse IgG (Dako, Copenhagen, Denmark) was used as the primary antibody. All slides were stained in one session. CD68-positive cells were enumerated at the posterior capsule including joint space and synovium [25 (link)].
Sogi Y., Yabe Y., Hagiwara Y., Tsuchiya M., Onoda Y., Sekiguchi T., Itaya N., Yoshida S., Yano T., Suzuki K., Onoki T, & Itoi E. (2020). Joint hemorrhage accelerates cartilage degeneration in a rat immobilized knee model. BMC Musculoskeletal Disorders, 21, 761.
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9

Immunohistochemical Staining of Tissue Sections

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Adjacent serial sections were routinely stained with H&E, immunohistochemically stained for SPARC (1:200 for 45 min; AF942, R&D Systems), the cell proliferation marker Ki-67 (1:250 for 40 min; M7249, Dako), and the macrophage/histiocyte marker CD68 (1:200 for 30 min; MCA341R, Serotec) as previously reported (36 (link)). Additional adjacent sections were subjected to the periodic acid Schiff (PAS) reaction with or without diastase (liver tissue was positive control) as previously described (43 (link)) to demonstrate phagocytic activity (histiocytes). Images of stained sections were captured on either a Nikon Eclipse E800 microscope with a Nikon DXM1200C digital camera using ImagePro 6.0 Plus (Media Cybernetics) software or a Nikon Eclipse Ni microscope with a Digital Sight DS-U3 digital camera using NIS-Elements AR 4.20 (Nikon) software. Composite figures were prepared using Adobe Photoshop CS6 software.
Thomas S.L., Schultz C.R., Mouzon E., Golembieski W.A., Naili R.E., Radakrishnan A., Lemke N., Poisson L.M., Gutiérrez J.A., Cottingham S, & Rempel S.A. (2015). Loss of Sparc in p 53‐null Astrocytes Promotes Macrophage Activation and Phagocytosis Resulting in Decreased Tumor Size and Tumor Cell Survival. Brain Pathology, 25(4), 391-400.
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10

Renal Tissue Immunohistochemistry Protocol

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Paraffin sections of renal tissue were cut at 4-μm thickness and subjected to microwave irradiation in citrate buffer to enhance antigen retrieval. After blocking steps with 0.3% hydrogen peroxide and non-fat milk, the following antibodies were used as primary antibodies: anti-rat CD68 (MCA341R, Serotec, Oxford, UK), anti-E-cadherin (Ecad, IS059; Dako Co, Denmark), anti-α-smooth muscle actin (αSMA; IS700; Dako Co, Denmark), anti-vimentin (M0725; Dako Co, Denmark), and anti-IDO (MAB5412; Merck Millipore, Billerica, MA). All antibodies were diluted 1:100. To complete the sandwich, sections were incubated with LSAB+ System-HRP reagents (K0690; Dako Co, Denmark). Finally, DAB substrate-chromogen was used to complete the reaction (K346811; Dako Co, Denmark).
We conducted a quantitative analysis of the positive interstitial cells for ED-1, αSMA and vimentin in a blinded fashion under X200 microscopic magnification, expressed as cells per field. The positive and negative tubular cells for αSMA and vimentin were counted under X200 microscopic magnification and the results are expressed as a percentage of positive cells. To analyze E-cadherin and IDO expression, the positive and negative tubules were counted for presenting as a percentage of positive tubules.
Matheus L.H., Simão G.M., Amaral T.A., Brito R.B., Malta C.S., Matos Y.S., Santana A.C., Rodrigues G.G., Albejante M.C., Bach E.E., Dalboni M.A., Camacho C.P, & Dellê H. (2017). Indoleamine 2, 3-dioxygenase (IDO) increases during renal fibrogenesis and its inhibition potentiates TGF-β 1-induced epithelial to mesenchymal transition. BMC Nephrology, 18, 287.
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