Cl8942ap

Manufactured by Cedarlane

Sourced in United States, Canada

The CL8942AP is a laboratory centrifuge designed for general-purpose applications. It features a compact design and can accommodate various sample sizes and volumes. The centrifuge provides reliable performance for routine sample preparation and separation tasks.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Image pro plus, by Media Cybernetics (2 mentions) Prolong gold antifade, by Thermo Fisher Scientific (2 mentions) Ab32457, by Cedarlane (2 mentions) A21206, by Abcam (2 mentions) D3571, by Thermo Fisher Scientific (2 mentions) Fluoresbrite polychromic red 1.0 micron beads, by Polysciences (2 mentions) Td 88137, by Inotiv (2 mentions) Ab28364, by Cedarlane (2 mentions) Perilipin, by Biosynth (2 mentions) Lsm 510 confocal microscope, by Zeiss (2 mentions) Ab13970, by Abcam (2 mentions) E0467, by Agilent Technologies (2 mentions) Anti mac2, by Cedarlane (2 mentions) Ab45371, by Abcam (1 mentions) F3777, by Merck Group (1 mentions) Fluoview fv1000 confocal microscope, by Olympus (1 mentions) Ab27478, by Abcam (1 mentions) Ab10558, by Abcam (1 mentions) A0545, by Merck Group (1 mentions)

37 protocols using cl8942ap

Cell-Specific TLR7 Expression in Atherosclerosis

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Cell type (macrophage, SMC, and EC)-specific expression of TLR7 in mouse atherosclerotic lesions was determined by immunofluorescent double staining using rabbit anti-mouse TLR7 monoclonal antibody (1:100, ab45371, Abcam) together with rat anti-mouse Mac-2 monoclonal antibody (macrophages, 1:100, CL8942AP, CEDARLANE, Burlington, NC), purified rat anti-mouse CD31 monoclonal antibody (EC, 1:900, #553370, BD Biosciences), and FITC-conjugated mouse anti-α-smooth muscle actin monoclonal antibody (SMC, 1:500, F3777, Sigma-Aldrich). Mouse aortic arch lesion macrophage and SMC apoptosis was also detected by immunofluorescent double staining with Alexa Fluor® 594-conjugate cleaved caspase-3 (Asp175) (D3E9) rabbit monoclonal antibody (1:200, #8172, Cell Signaling Technology, Danvers, MA) together with rat anti-mouse Mac-2 monoclonal antibody or FITC-conjugated mouse anti-α-smooth muscle actin monoclonal antibody as mentioned above. Images were collected under an Olympus FluoView™ FV1000 Confocal Microscope.

Liu C.L., Santos M.M., Fernandes C., Liao M., Iamarene K., Zhang J.Y., Sukhova G.K, & Shi G.P. (2017). Toll-like receptor 7 deficiency protects apolipoprotein E-deficient mice from diet-induced atherosclerosis. Scientific Reports, 7, 847.

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Colon Tissue Immunohistochemistry Protocol

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Paraffin-embedded colon tissue were sectioned at 5 μm, deparaffinized and permeabilized with Triton X-100 (Sigma-Aldrich) and heat antigen retrieved with Bull’s Eye Decloaker (Biocare Medical). Slides were then blocked for endogenous peroxidase with Bloxall^™ blocking solution (Vector Lab) followed by blocking with either 3% bovine serum albumin, or mouse on mouse blocking reagent (Vector Lab). Anti-IL11 (ab10558; PA5-36544, Invitrogen), anti-CD45 (1:100; ab10558, Abcam), anti-LGALS3 (1 μg/ml; CL8942AP, Cedarlane) amd anti-LAMP2 (1 μg/ml; 550292, BD Bioscience) were added and incubated overnight at 4°C. Anti-rabbit (1:100; ab27478, Abcam) and anti-rat IgG (1 μg/ml; sc-2026, Santa Cruz) isotype controls were added as respective negative controls. Slides were incubated with anti-rabbit IgG peroxidase (1:500, A0545, Sigma-Aldrich) and anti-rat IgG peroxidase (MP-7404, Vector Lab) followed by chromogen development with ImmPACT^® DAB peroxidase substrate kit (SK-4105, Vector Lab) according to manufacturer’s instructions. Lastly, Gill’s haematoxylin (H-3401, Vector Lab) was added for nuclear counterstain. To control for unspecific binding, primary antibody isotype controls were included and images are presented in S3 Fig.

Lim W.W., Ng B., Widjaja A., Xie C., Su L., Ko N., Lim S.Y., Kwek X.Y., Lim S., Cook S.A, & Schafer S. (2020). Transgenic interleukin 11 expression causes cross-tissue fibro-inflammation and an inflammatory bowel phenotype in mice. PLoS ONE, 15(1), e0227505.

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Immunohistological Analysis of Adipocyte Size

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For immunohistology, tissues were fixed with neutral buffered 10% formalin solution, embedded in paraffin, cut into sections, and then deparaffinized. Sections were incubated with anti-Mac-2 (CL8942AP, Cedarlane Labs) for 30 min at room temperature, and counterstained with hematoxylin. Adipocyte size was measured on 5 independent H&E-stained sections per animal. Oil Red O (ORO) staining was performed on OCT-embedded sections. Images were captured by a digital system, and the staining area was measured using computer-assisted image quantification (Image-Pro Plus software, Media Cybernetics).

Wara A.K., Wang S., Wu C., Fang F., Haemmig S., Weber B.N., Aydogan C.O., Tesmenitsky Y., Aliakbarian H., Hawse J.R., Subramaniam M., Zhao L., Sage P.T., Tavakkoli A., Garza A., Lynch L., Banks A.S, & Feinberg M.W. (2020). KLF10 Deficiency in CD4+ T Cells Triggers Obesity, Insulin Resistance, and Fatty Liver. Cell reports, 33(13), 108550.

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Characterizing Smad7 in Infarct Macrophages

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In order to systematically characterize the time course of Smad7 expression in infarct macrophages, Smad7 (using an anti-Smad7 antibody by Thermo Fisher Scientific, #42-0400) and GFP staining (using an anti-GFP antibody, Abcam, #ab6662) was performed on the infarcted CSF1R^EGFP heart at baseline, 24h, 3-day, 7-day and 28-day timepoints (n=5-6/each time point). In order to identify myofibroblasts in the infarct, sections were stained with an anti-α-smooth muscle actin (SMA) antibody (Sigma, #F3777) as previously described⁴¹. Myofibroblasts were identified as spindle-shaped α-SMA-positive cells located outside the vascular media. To determine the number of macrophages in the infarcted and remodeling myocardium, sections from infarcted Smad7 fl/fl and MyS7KO mice were stained with an anti-Mac2 (galectin-3) antibody (Cedarlane, #CL8942AP), as previously described^{42 (link),43 (link)}. Quantitative analysis was performed by counting the number of macrophages in 10 fields from 2 different levels for each animal.

Li J., Li R., Tuleta I., Hernandez S.C., Humeres C., Hanna A., Chen B, & Frangogiannis N.G. (2022). The role of endogenous Smad7 in regulating macrophage phenotype following myocardial infarction. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 36(7), e22400.

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Immunofluorescence Staining of Mouse Heart Sections

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Five micron sections of paraffin-embedded mouse hearts were deparaffinised in xylene, rehydrated in alcohol series and immersed in distilled water, followed by high-temperature antigen retrieval in citrate buffer (pH 6) and blocked with 0.5% bovine serum albumin (BSA). For immunofluorescence, the slides were stained with primary antibodies against CCR7 (NB110-55680, Novus Biologicals, Littleton, CO; 1∶100), CD31 (DIA-310, Dianova GmbH, Hamburg, Germany; 1∶100), Fibroblast Marker (sc-73355, Santa Cruz Biotechnology, San Diego, CA; 1∶100), α smooth muscle Actin (ab7817, abcam, Cambridge, UK; 1∶100) or Mac2 (CL8942AP, Cedarlane, Burlington, Canada; 1∶400) for 1 hour at room temperature and counterstained with Alexa Fluor 488 goat anti-rabbit IgG (1∶500), Alexa Fluor 568 goat anti-rat IgG (1∶500), Alexa Fluor 568 goat anti-mouse or Rat on Mouse AP Polymer kit in combination with Warp Red Chromogen kit (Biocare Medical, San Francisco, CA), respectively. The slides were mounted with SlowFade Gold antifade reagent with DAPI (Life Technologies, Carlsbad, CA). Images were taken by a Nikon Eclipse E400 microscope (Tokyo, Japan).

Finsen A.V., Ueland T., Sjaastad I., Ranheim T., Ahmed M.S., Dahl C.P., Askevold E.T., Aakhus S., Husberg C., Fiane A.E., Lipp M., Gullestad L., Christensen G., Aukrust P, & Yndestad A. (2014). The Homeostatic Chemokine CCL21 Predicts Mortality in Aortic Stenosis Patients and Modulates Left Ventricular Remodeling. PLoS ONE, 9(11), e112172.

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Multicolor Immunofluorescence Staining Protocol

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After blocking, slides were incubated with the following primary antibodies: CD31 (Abcam ab32457, 1:100), LGALS3 (Cedarlane CL8942AP, 1:500) or their isotype IgG controls. Each slide had at least two sections stained with primary antibody and at least one section used as an IgG control. ACTA2 was conjugated to -FITC (Sigma F3777 clone 1A4 1:500). Secondary antibodies for immunofluorescence were as follows: donkey anti-rabbit 555 (Invitrogen A21206, 1:250) and donkey anti-rat Dylight 650 (Abcam ab102263, 1:250). Nuclear counterstain was performed with DAPI (ThermoFisher Scientific D3571), and slides were mounted with Prolong Gold Antifade (Invitrogen).

Newman A.A., Serbulea V., Baylis R.A., Shankman L.S., Bradley X., Alencar G.F., Owsiany K., Deaton R.A., Karnewar S., Shamsuzzaman S., Salamon A., Reddy M.S., Guo L., Finn A., Virmani R., Cherepanova O.A, & Owens G.K. (2021). Multiple cell types contribute to the atherosclerotic lesion fibrous cap by PDGFRβ and bioenergetic mechanisms. Nature metabolism, 3(2), 166-181.

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Atherosclerosis Plaque Imaging in Mice

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Apoe^−/− Myh11 Cre ER^T2 R26R-YFP mice fed a Western Diet (Harlan Teklad; TD.88137) for 17 weeks received an injection of Fluoresbrite Polychromic Red 1.0 micron beads (2.5% solids-latex; Polysciences, Inc., #18660). Beads were diluted in sterile PBS (1:4 dilution) and 250 μL of diluted beads were injected retro-orbitally. 24 hours after the injection, blood was collected by tail bleeding and selective uptake of the beads by circulating Ly6C^low monocytes was assessed by flow cytometry (see Flow cytometry section). 5–7 days after the bead injection, we started the anti-IL-1β antibody or IgG control regimen (10 mg/kg) for a duration of 3 weeks as described above. At the end of the antibody treatment, mice were euthanized by asphyxiation. Mice were perfusion fixed with Periodate-Lysine-Paraformaldehyde (PLP). Each BCA was harvested, post-fixed in PLP, and incubated in a gradient of sucrose solutions before being embedded in O.C.T. compound. Serial frozen 10 μm-thick sections were used to perform immunofluorescent staining for LGALS3 (Cedarlane CL8942AP) and DAPI for nuclear staining. Endogenous fluorescent signal of Fluoresbrite Polychromic Red beads was directly detectable.

Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.

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Histological Analysis of Fat and Liver

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Fat pads and livers were excised and fixed overnight in 10% PBS buffered formalin and were thereafter stored in 50% ethanol. Tissues were sectioned (5um) rehydrated and stained using H+E, or with primary antibodies to PTEN (Abcam 32199; Dilution 1:25) or Mac-2 (CL8942AP; Dilution 1:200, CEDARLANE Laboratories USA Inc.). Slides were mounted using Prolong Gold Antifade reagent with DAPI (Life Technologies).

Morley T.S., Xia J.Y, & Scherer P.E. (2015). Selective enhancement of insulin sensitivity in the mature adipocyte is sufficient for systemic metabolic improvements. Nature Communications, 6, 7906.

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Immunohistochemical Analysis of Invasive Breast Cancer

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Patient slices were obtained from the National Disease Research Interchange. All patients were diagnosed with invasive lobular/ductal carcinoma breast, Grade 2. Patients received total mastectomy or simple radical mastectomy with no known presurgical therapy. Samples were analyzed under the approved University of Texas Southwestern Medical Center Institutional Review Board protocol # STU 012017-043 (Endotrophin Upregulation in Tumor Tissues). Mouse tissues were dissected and fixed in 10% formalin overnight. Paraffin processing, embedding, sectioning and hematoxylin/eosin staining and trichrome staining were performed by John Sheldon at UTSW Medical Center. Immunostaining was performed following standard protocols, with the following antibodies: Perilipin (1:1000, #20R-PP004, Fitzgerald), EGFP (1:500, ab13970, Abcam), α-SMA (1:500, eBioscience™, #14-9760-80), CD31 (1:200, Abcam, #ab28364), Mac2 (1:750, CL8942AP, CEDARLANE) and collagen hybridizing peptide (#5276, Advanced BioMatrix). Images were acquired with LSM510 confocal microscope (Zeiss) and analyzed by Image J software.

Zhu Q., Zhu Y., Hepler C., Zhang Q., Park J., Gliniak C., Henry G.H., Crewe C., Bu D., Zhang Z., Zhao S., Morley T., Li N., Kim D.S., Strand D., Deng Y., Robino J.J., Varlamov O., Gordillo R., Kolonin M.G., Kusminski C.M., Gupta R.K, & Scherer P.E. (2022). Adipocyte mesenchymal transition contributes to mammary tumor progression. Cell reports, 40(11), 111362.

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Macrophage Immunohistochemistry Protocol

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Tissues were fixed for 12–24 h in 10% formalin, embedded in paraffin, and sectioned. Sections were incubated overnight at 4°C with rat anti-Mac2 (CL8942AP; Cedarlane). Biotinylated rabbit anti-rat (E0467; Dako) was used as secondary antibody. The ABC peroxidase kit (Pierce) was used for immunodetection, and sections were counterstained in Mayer’s hematoxylin.

Casana E., Jimenez V., Jambrina C., Sacristan V., Muñoz S., Rodo J., Grass I., Garcia M., Mallol C., León X., Casellas A., Sánchez V., Franckhauser S., Ferré T., Marcó S, & Bosch F. (2022). AAV-mediated BMP7 gene therapy counteracts insulin resistance and obesity. Molecular Therapy. Methods & Clinical Development, 25, 190-204.

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