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M11217

Manufactured by Merck Group
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The M11217 is a laboratory centrifuge designed for general laboratory applications. It is capable of separating various samples through high-speed rotation. The centrifuge provides a means of isolating and concentrating specific components within a liquid or solid mixture.

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Lab products found in correlation

Anti mcherry, by Thermo Fisher Scientific (3 mentions) H3663, by Santa Cruz Biotechnology (2 mentions) Sc 56, by Abcam (2 mentions) Anti h4k20me2 3, by Abcam (2 mentions) Alexa fluor conjugated series, by Thermo Fisher Scientific (2 mentions) Lsm 700 multiphoton confocal microscope, by Adobe (2 mentions) Anti h3k27me3, by Merck Group (2 mentions) Lsm 880, by Zeiss (2 mentions) Goat anti mouse alexa633, by Thermo Fisher Scientific (1 mentions) Neurotrace 435 455, by Thermo Fisher Scientific (1 mentions) Alexa 594 donkey anti rat, by Thermo Fisher Scientific (1 mentions) Mab1572, by Thermo Fisher Scientific (1 mentions) Ab192761, by Abcam (1 mentions) Ab76020, by Abcam (1 mentions) A2228, by Merck Group (1 mentions) Apc 012, by Alomone (1 mentions) Mms 101p, by BioLegend (1 mentions) Alexa fluor labeled secondary antibody, by Thermo Fisher Scientific (1 mentions) Ab980, by Merck Group (1 mentions) S5545, by Merck Group (1 mentions)

6 protocols using m11217

1

Immunofluorescence Staining of Cellular Markers

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Immunofluorescence staining was performed using standard procedures7 (link),71 . Primary antibodies used were mouse anti-Fas III (1:200, DSHB, 7G10), anti-HA (1:200; Sigma-Aldrich H3663), anti-PCNA (1:200; Santa Cruz sc-56), anti-GFP (1:1,000; Abcam ab 13970), anti-mKO (1:200; MBL PM051M), anti-mCherry (1:1000; Invitrogen M11217), anti-H3K27me3 (1:200; Millipore 07–449), anti-H4K20me2/3 (1:400; Abcam ab7817), anti-ssDNA (1:100, DSHB) and anti-BrdU (1:200; Abcam ab6326). BrdU analogue was Invitrogen B23151 5-bromo-2´-deoxyuridine (BrdU). Secondary antibodies were the Alexa Fluor-conjugated series (1:1000; Molecular Probes). Confocal images for immunostained fixed sample were taken using Zeiss LSM 700 Multiphoton confocal microscope with 63x or 100x oil immersion objectives and processed using Adobe Photoshop software.
Wooten M., Snedeker J., Nizami Z.F., Yang X., Ranjan R., Urban E., Kim J.M., Gall J., Xiao J, & Chen X. (2019). Asymmetric histone inheritance via strand-specific incorporation and biased replication fork movement. Nature structural & molecular biology, 26(8), 732-743.
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2

Immunofluorescence Staining of Cellular Markers

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Immunofluorescence staining was performed using standard procedures7 (link),71 . Primary antibodies used were mouse anti-Fas III (1:200, DSHB, 7G10), anti-HA (1:200; Sigma-Aldrich H3663), anti-PCNA (1:200; Santa Cruz sc-56), anti-GFP (1:1,000; Abcam ab 13970), anti-mKO (1:200; MBL PM051M), anti-mCherry (1:1000; Invitrogen M11217), anti-H3K27me3 (1:200; Millipore 07–449), anti-H4K20me2/3 (1:400; Abcam ab7817), anti-ssDNA (1:100, DSHB) and anti-BrdU (1:200; Abcam ab6326). BrdU analogue was Invitrogen B23151 5-bromo-2´-deoxyuridine (BrdU). Secondary antibodies were the Alexa Fluor-conjugated series (1:1000; Molecular Probes). Confocal images for immunostained fixed sample were taken using Zeiss LSM 700 Multiphoton confocal microscope with 63x or 100x oil immersion objectives and processed using Adobe Photoshop software.
Wooten M., Snedeker J., Nizami Z.F., Yang X., Ranjan R., Urban E., Kim J.M., Gall J., Xiao J, & Chen X. (2019). Asymmetric histone inheritance via strand-specific incorporation and biased replication fork movement. Nature structural & molecular biology, 26(8), 732-743.
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3

Immunohistochemistry of Labeled Neurons

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After slicing, samples for immunohistochemistry were blocked with 2.5% normal donkey serum / 2.5% normal goat serum / 0.5% Triton X-100/PBS for 4–6 hours at room temperature, primary antibodies for 4–6 days at 4°C, and secondary antibodies overnight at 4°C. Antibodies were diluted in blocking solution. The following antibodies were used: rat anti-mCherry (1:250, ThermoFisher M11217), Mouse anti-Parvalbumin (1:1000, Millipore MAB1572), Donkey anti-Rat-Alexa 594 (1:1000, Invitrogen), and Goat anti-Mouse-Alexa 633 (1:1000, Invitrogen). Neurotrace 435/455 (1:250, ThermoFisher N21479) was added to the secondary antibody solution. Sections were mounted and imaged on a Zeiss LSM 880 using a 20x objective in 425×425 μm tiles at 1024×1024-pixel identification.
Beau M., Herzfeld D.J., Naveros F., Hemelt M.E., D’Agostino F., Oostland M., Sánchez-López A., Chung Y.Y., Michael M.a.i.b.a.c.h., Kyranakis S., Stabb H.N., Martínez Lopera M.G., Lajko A., Zedler M., Ohmae S., Hall N.J., Clark B.A., Cohen D., Lisberger S.G., Kostadinov D., Hull C., Häusser M, & Medina J.F. (2024). A deep-learning strategy to identify cell types across species from high-density extracellular recordings. bioRxiv.
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4

Expression and Characterization of KCNG4 Mutant

Cited 1 time
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A full-length human KCNG4 cDNA clone was purchased from Source bioscience (IRCMp5012B0629D) and cloned in-house into a pcDNA3 based expression plasmid (CMV-genex-polioIRESmCherry) both with or without a C-terminal HA tag. The p.Val419Met mutation was introduced by site-directed mutagenesis (Stratagene) according to the manufacturer’s protocols and sequences of the plasmids were confirmed by Sanger Sequencing. The clone expressing KV2.1 alongside a nuclear GFP reporter in the pCAGGS-IRES2-nucEGFP vector has been described previously (Saitsu et al., 2015 (link)) and was a kind gift from Prof. Hiromoto Saitsu.
Antibodies used were anti-HA mouse monoclonal (12B12, #MMS-101P, Biolegend), anti-Na+/K+ ATPase rabbit monoclonal (ab76020, Abcam), anti-mCherry rat monoclonal (M11217), anti-β-actin mouse monoclonal (a2228, Sigma), anti-KV2.1 rabbit polyclonal (APC-012, Alomone), anti-KV6.4 mouse monoclonal (N458/10, NeuroMab), anti-KV2.1 mouse monoclonal (K89/34, ab192761, Abcam), and anti-HA rabbit monoclonal (C29F4 #3724, Cell Signaling).
Lee M.C., Nahorski M.S., Hockley J.R., Lu V.B., Ison G., Pattison L.A., Callejo G., Stouffer K., Fletcher E., Brown C., Drissi I., Wheeler D., Ernfors P., Menon D., Reimann F., Smith E.S, & Woods C.G. (2020). Human Labor Pain Is Influenced by the Voltage-Gated Potassium Channel KV6.4 Subunit. Cell Reports, 32(3), 107941.
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5

RGC-Oligodendrocyte Co-culture Myelination

Cited 2 times
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Retinae were dissected from P6 Sprague-Dawley rat pups (AMPREP Animal Ethics Committee #E-1602/2015/M). RGCs were purified according to the published immunopanning protocol (Watkins et al., 2008 (link); Deliyanti and Wilkinson-Berka, 2015 (link)). Details of the RGC preparation are described in Supplemental Experimental Procedure. The RGC growth medium was changed every third day and cultures were maintained for 9 days. Pre-OLs from NKX2.1-GFP + cells at stage VI day 10, were FACS sorted according to O4 antibody labeling. O4 + sorted cells were then co-cultured with RGCs at a density of 20,000/well under 3 different conditions: T3 with 0.01% ethanol, 10 ng/mL DITPA, and 40 ng/mL T3 with DITPA in myelination medium (see Supplemental Experimental Procedure). The medium was changed every 3 days. At day 7, all co-cultures were fixed with 2% paraformaldehyde (PFA) for 10 min at room temperature. Cells were stained with a monoclonal rat anti-mCherry (Life Technologies, M11217, 1:1000); monoclonal mouse anti-NF-200 (Sigma-Aldrich, N0142, 1:200); and polyclonal rabbit anti-MBP (Millipore, AB980, 1:200). Subsequently cells were stained with appropriate Alexa Fluor-labeled secondary antibodies (Life technologies).
Lee J.Y., Kim M.J., Deliyanti D., Azari M.F., Rossello F., Costin A., Ramm G., Stanley E.G., Elefanty A.G., Wilkinson-Berka J.L, & Petratos S. (2017). Overcoming Monocarboxylate Transporter 8 (MCT8)-Deficiency to Promote Human Oligodendrocyte Differentiation and Myelination. EBioMedicine, 25, 122-135.
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6

Viral Expression Visualization in Mouse Brain

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To verify viral expression, mice were deeply anaesthetized with isoflurane and transcardially perfused with PBS, followed by fixation in cold 4% PFA solution for 48 h. Then the brain tissue was dissected and fixed in PFA overnight at 4 °C. The brain tissue was dehydrated in PBS solution containing 30% sucrose and then embedded in OCT freezing embedding solution. Coronal sections (30 μm) were cut on a Leica CM1860 UV freezing microtome (20 sections per brain). The brain slices were permeabilized in PBST for 20 min, subsequently blocked with 5% BSA in PBST for 1 h at room temperature, and then incubated with anti-mCherry (Thermo Fisher M11217,1:1000) and anti-Serotonin (Sigma–Aldrich s5545,1:2000) overnight at 4 °C. After washing with PBS for 10 min and PBST twice for 10 min each, brain slices were incubated in Alexa Fluor 568 Goat anti-Rat (Thermo Fisher, 1:400) and Alexa Fluor 488 Donkey anti-Rabbit (Jackson Labs, 1:200) for 3 h at room temperature. After washing 3 times with PBS for 10 min each, brain slices were sealed with anti-fading fluorescent mounting medium (Vectorlabs) overlay and subsequently imaged with a confocal microscope.
Zhao Y., Huang C.X., Gu Y., Zhao Y., Ren W., Wang Y., Chen J., Guan N.N, & Song J. (2024). Serotonergic modulation of vigilance states in zebrafish and mice. Nature Communications, 15, 2596.
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