Af2864

Manufactured by Merck Group

AF2864 is a laboratory equipment item produced by Merck Group. It is designed for general laboratory use, but a detailed description is not available at this time.

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Lab products found in correlation

Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Alexa 647, by Thermo Fisher Scientific (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) Ab9348, by Merck Group (1 mentions) Ab5888, by Merck Group (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) Nanozoomer s60, by Hamamatsu Photonics (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidine 2 phenylindole dihydrochloride dapi, by Merck Group (1 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions) Olig2, by Merck Group (1 mentions) Ab21624, by R&D Systems (1 mentions) Ab9610, by Proteintech (1 mentions) Sox10, by R&D Systems (1 mentions) Mab1195, by R&D Systems (1 mentions) Prb 278p, by Santa Cruz Biotechnology (1 mentions) Smi 99p, by Fortrea (1 mentions) Ab9610, by Merck Group (1 mentions) Mab353, by Merck Group (1 mentions)

4 protocols using af2864

Immunohistochemical Analysis of MS Brain Tissue

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Autopsy brain tissue samples from patients with confirmed secondary
progressive MS were obtained from the United Kingdom MS tissue bank
(Richards Reynolds, Imperial College, London). MS tissue block containing
active lesions and periplaque white matter were selected for analysis.
Cryosections (14 μm thick) of snap frozen MS brain tissue were
permeabilized and blocked in blocking buffer (0.05% Triton X-100 and
10% normal goat serum in PBS) for 1h and overlaid with primary
antibodies overnight at 4 °C. Antibodies used in the study were:
rabbit anti-CHD8 (Bethyl, A301-225A, 1:1000), goat anti-Sox10 (R&D
systems, AF2864, 1:400), rabbit anti-Nogo A (Millipore, AB5888, 1:200),
chicken anti-MBP (Millipore, AB9348, 1/50). After washing with 0.05%
Triton X-100 in PBS, sections were incubated with secondary antibodies
conjugated to Alexa488, Alexa594 or Alexa647 (Thermo, 1:1,000) and DAPI for
1h at room temperature, washed in PBS and mounted with Fluoromount-G
(SouthernBiotech). Pictures of were taken with Zeiss microscope using
apotome. Z-stack was used to average 5–7 planes and pictures were
treated and cells were counted using Zen and imageJ software packages.

Zhao C., Dong C., Frah M., Deng Y., Marie C., Zhang F., Xu L., Ma Z., Dong X., Lin Y., Koenig S., Nait-Oumesmar B., Martin D.M., Wu L.N., Xin M., Zhou W., Parras C, & Lu Q.R. (2018). Dual-requirement of CHD8 for Chromatin Landscape Establishment and Histone Methyltransferase Recruitment to Promote CNS Myelination and Myelin Repair. Developmental cell, 45(6), 753-768.e8.

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Quaternary Ammonium and Organophosphate Impacts on Cortical Organoids

Cited 1 time

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Cortical organoids were treated on alternating days between days 61 to 70 with quaternary ammonium and phosphonium compounds or organophosphate flame retardants at their approximate IC₉₀ and IC₇₅ concentrations respectively. Organoids were harvested on day 70, washed in PBS, and fixed overnight with ice-cold 4% Paraformaldehyde (Electron Microscopy Sciences, HP1–100Kit). On the following day organoids were washed with PBS and cryoprotected using a 30% sucrose solution. Organoids were then embedded in OCT and sectioned at 15 μM. Slides were washed with PBS and incubated overnight with anti-SOX10 (1:200, R&D, AF2864) and anti-APC CC1 (1:200, Millipore, MABC200), followed by labeling with Alexa Fluor-conjugated secondary antibodies (2 μg/mL, Thermo Fisher). Slides were imaged at 10x magnification using a Hamamatsu Nanozoomer S60. Quantification of positive cells was performed using QuPath software (https://qupath.github.io/)^{52 (link)}.

Cohn E.F., Clayton B.L., Madhavan M., Yacoub S., Federov Y., Paul-Friedman K., Shafer T.J, & Tesar P.J. (2023). Pervasive environmental chemicals impair oligodendrocyte development. bioRxiv.

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Immunostaining Protocol for Cell Characterization

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Cells were prepared for immunostaining by fixation in 4% PFA (Electron Microscopy Sciences) for 15 min and subsequent permeabilization for 10 min with 0.2% Triton X-100 in 1× PBS. Cells were then blocked for non-specific binding with filtered 10% normal donkey serum (Abcam) in 1× PBS for 1 h at room temperature. Primary antibodies were diluted in blocking solution and incubated with the samples overnight at 4 °C. Samples were rinsed with 1× PBS and incubated with the appropriate fluorescently labeled Alexa-Fluor secondary antibodies (Thermo Fisher, 1:500) for 1 h at room temperature. For nuclear staining, samples were incubated with 1 μg/ml 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) (Sigma) for 5 min. Primary antibodies used were: Sox10 (R&D Systems, AF2864; 2 µg/ml), Olig2 (Millipore, AB9610 or ProteinTech, 13999-1-AP; 1:1000), Sox1 (R&D Systems, AF3369; 1 µg/ml), Pax6 (Covance, PRB-278P; 0.67 µg/ml), Oct4 (Santa Cruz, SC-5279; 0.4 µg/ml), Nkx2.2 (DSHB, 74.5A5; 4.4 μg/ml), MBP (Covance, SMI-99P; 2 µg/ml), O4 (kindly gifted from Robert Miller; 1:10), PLP1 (kindly gifted from Bruce Trapp; 1:200), Nanog (Abcam, AB21624; 1 µg/ml), and βIII-Tubulin (R&D Systems, MAB1195; 1 µg/ml).

Lager A.M., Corradin O.G., Cregg J.M., Elitt M.S., Shick H.E., Clayton B.L., Allan K.C., Olsen H.E., Madhavan M, & Tesar P.J. (2018). Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells. Nature Communications, 9, 3708.

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Immunostaining and in situ Hybridization of Brain Sections

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Antibodies and their dilutions used for immunostaining of brain sections: goat anti-PDGFRα (R&D Systems AF1062, 1:50); rabbit anti-PDGFRα (Santa Cruz Biotechnology SC-338, 1:50); rabbit anti-Ki67 (Abcam 15580, 1:500); rabbit anti-Olig2 (Millipore AB9610, 1:500); rabbit anti-Sox2 (Millipore AB5603, 1:200); goat anti-Sox10 (R&D Systems AF2864, 1:50); mouse anti-Nestin (Millipore MAB353, 1:250); rabbit anti-Ng2 (Millipore AB520, 1:500); rabbit anti-NeuN (Novus NBP1-77686SS, 1:200); rabbit anti-GFAP (Invitrogen 180063, 1:200); mouse anti-APC, clone CC1 (Calbiochem OP80, 1:20).
Secondary antibodies were coupled with Alexa fluorophores (Jackson Immunoresearch) or biotinylated for ABC signal amplification and peroxidase DAB staining (Vector Labs).
For mRNA in situ hybridization, cryosections were hybridized with Digoxigenin-labeled riboprobes, which were synthesised from the plasmids pBS-Pdgfra-ISH-1.6 kb and pCR4-Plp1-ISH-2.0 kb, and probe hybridization was detected with AP-conjugated anti-Dig Fab fragments (Roche) and NBT/BCIP staining.
Hematoxylin & Eosin staining was performed with Shandon Gill #2 Hematoxylin and Eosin-Y (Thermo Scientific).

Zou H., Feng R., Huang Y., Tripodi J., Najfeld V., Tsankova N.M., Jahanshahi M., Olson L.E., Soriano P, & Friedel R.H. (2015). Double minute amplification of mutant PDGF receptor α in a mouse glioma model. Scientific Reports, 5, 8468.

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