Ma1 81510

Manufactured by Thermo Fisher Scientific

The MA1-81510 is a laboratory instrument designed for general analytical applications. It provides accurate and reliable measurements, but a detailed description of its core function cannot be provided in a concise, unbiased, and factual manner without the risk of extrapolation or interpretation.

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Lab products found in correlation

C9205, by Merck Group (2 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Ab157107, by Abcam (1 mentions) Ab109186, by Abcam (1 mentions) Ab33786, by Abcam (1 mentions) Ab22858, by Abcam (1 mentions) Ab5076, by Abcam (1 mentions) Mab1435, by Merck Group (1 mentions) Ab5690, by Abcam (1 mentions) Eclipse 90i, by Nikon (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Gtx102090, by GeneTex (1 mentions) Nis elements ar software, by Nikon (1 mentions) Mab377, by Merck Group (1 mentions) Goat anti tnf α, by Santa Cruz Biotechnology (1 mentions) Sc 1506, by Santa Cruz Biotechnology (1 mentions) Sc 1350, by Santa Cruz Biotechnology (1 mentions)

4 protocols using ma1 81510

Immunohistochemical Analysis of Rat Brain Tumor Microenvironment

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Coronal cryosections (12 μm) of rat brains were obtained from the center of the anterior-posterior extent of the tumor. Tissues were blocked for 1 h in 2% donkey serum with 0.2% Triton X-100 and labeled overnight at 4°C with primary antibodies against GFAP (1:500; C9205, Sigma-Aldrich), ED-1 (1:200; MAB 1435, Millipore, Billerica, MA), RECA-1 (1:200; MA1-81510, Thermo Fisher Scientific Inc.), HIF-1 (1:200; sc-12542, Santa Cruz Biotechnology, Inc.), CD44 (1:200; ab157107, Abcam), Olig2 (1:100; ab109186, Abcam), CD3 (1:200; ab5690, Abcam), CD8a (1:200; ab33786, Abcam), CD4 (1:200; sc-1573, Santa Cruz Biotechnology, Inc, CD49d (1:200; ab22858, Abcam), P2Y12 (1:200; AS-55043, AnaSpec, Fremont, CA) or IBA1 (1:100; ab5076, Abcam). Alexa Fluor 500- or fluorescein isothiocyanate-conjugated secondary antibodies were applied, and tissues were coverslipped with ProLong Gold antifade reagent (P36930; Thermo Fisher Scientific Inc.). Specific labeling was confirmed by omission of primary antibody. Immunolabeled tissues were visualized with epifluorescence microscopy (Nikon Eclipse 90i; Nikon Instruments Inc., Melville, NY).

Connolly N.P., Stokum J.A., Schneider C.S., Ozawa T., Xu S., Galisteo R., Castellani R.J., Kim A.J., Simard J.M., Winkles J.A., Holland E.C, & Woodworth G.F. (2017). Genetically engineered rat gliomas: PDGF-driven tumor initiation and progression in tv-a transgenic rats recreate key features of human brain cancer. PLoS ONE, 12(3), e0174557.

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Endothelial Cell Survival Visualization

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Hoechst/RECA-1 was used to visualize the survival of ECs [39 (link)]. Analysis was facilitated by the use of a fluorescent nuclear dye, Hoechst 33342 (5 μM). Slides were stained with RECA-1 antibody (rat endothelial cell antigen-I, MA1-81510; Thermo Fisher Scientific Inc.) to visualize endothelial cells. RECA-1 was used at a dilution of 150 and was revealed using donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor™ 594 (Thermo Fisher Scientific Inc., 1:500). For quantification of the area of Hoechst⁺ and RECA-1⁺ endothelial cells, NIH ImageJ software was utilized. The results were displayed as number of positive cells per micrometer.

Chen Z., Wang X., Wu H., Fan Y., Yan Z., Lu C., Ouyang H., Zhang S, & Zhang M. (2022). X-box binding protein 1 as a key modulator in “healing endothelial cells”, a novel EC phenotype promoting angiogenesis after MCAO. Cellular & Molecular Biology Letters, 27, 97.

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Immunohistochemical Analysis of Nuclear Factor-κB Activation in the ACA-MCA Watershed Territory

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Coronal cryosections (12 μm) were blocked for 1 hour in 2% donkey serum and 0.2% TritonX-100. Sections were incubated overnight at 4 °C with primary antibody against p65 (GTX102090; Genetex, Irvine CA) and double-labeled with primary antibody against rat endothelial cell antigen-1 (RECA-1) (MA1-81510; Thermo Fisher Scientific Inc.). Species appropriate Alexa Fluor 555- or fluorescein isothiocyanate-conjugated secondary antibodies were used for visualization. Sections were coverslipped with ProLong Gold antifade reagent (P36930; Thermo Fisher Scientific Inc.) containing the nuclear stain, DAPI (4′,6-Diamidine-2′-phenylindole dihydrochloride). Omission of primary antibody was used as a negative control.
Unbiased measurements of signal intensity within regions of interest (ROIs) were obtained using NIS-Elements AR software (Nikon Instruments, Melville, NY, USA), as described previously [28 (link)]. The area that was evaluated was a rectangle, 360 μm × 440 μm, positioned within the ACA-MCA watershed territory. Specific labeling for p65 within the ROI was defined as pixels with signal intensity >2.0× background. For nuclear translocation of p65, the ROI was defined by DAPI labeling within the same territory.

Gerzanich V., Kwon M.S., Woo S.K., Ivanov A, & Simard J.M. (2018). SUR1-TRPM4 channel activation and phasic secretion of MMP-9 induced by tPA in brain endothelial cells. PLoS ONE, 13(4), e0195526.

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Antibody Characterization for Neurological Study

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The custom anti-Trpm4 and anti-Sur1 antibodies used for this study have been previously described (15 (link)). The antigenic peptide for Trpm4 was the N-terminal intracellular domain of mouse Trpm4, corresponding to amino acids 1–612 (NP 780339). Anti-Trpm4 antibodies were raised in chicken, and were used at 1:200 dilution. The antigenic peptide for Sur1 was the intra-cellular nucleotide-binding domain 1 of Sur1 (rat Sur1 cDNA amino acids 598 to 965 of NP_037171). Anti-Sur1 antibodies were raised in rabbit and were used at 1:200 dilution. Other primary antibodies included: rabbit anti-cytokeratin 20 (prediluted; Ventana, Tucson, AZ) for colonic epithelial cells; mouse anti-glial fibrillary acidic protein (GFAP) (1:500; CY3 conjugated; C-9205; Sigma, St. Louis, MO) for astrocytes; mouse anti-NeuN (1:100; MAB377; Chemicon, Temecula, CA) for neurons; goat anti-CD31 (PECAM-1) (1:200; sc-1506; Santa Cruz Biotechnology, Santa Cruz, CA) for endothelial cells; goat anti-IgG (1:500; FITC conjugated; NB7477; Novus Biologicals, Littleton, CO) for immunoglobulin G (IgG); goat anti-TNFα (1:100; sc-1350; Santa Cruz Biotechnology, Santa Cruz, CA) for tumor necrosis factor α; rabbit anti-myeloperoxidase (MPO) (1:200, A0398; Dako, Carpinteria, CA) for neutrophils; and mouse anti-rat endothelial cell antigen (Reca-1) (1:100; MA1-81510; Thermo Fisher, Rockford, IL) for rat endothelium.

Mehta R.I., Tosun C., Ivanova S., Tsymbalyuk N., Famakin B.M., Kwon M.S., Castellani R.J., Gerzanich V, & Simard J.M. (2015). Sur1-Trpm4 Cation Channel Expression in Human Cerebral Infarcts. Journal of neuropathology and experimental neurology, 74(8), 835-849.

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