A 21311

Manufactured by Thermo Fisher Scientific

Sourced in United States

The A-21311 is a high-performance laboratory centrifuge designed for various applications. It features a microprocessor-controlled system that allows for precise speed and time settings. The centrifuge can accommodate a range of rotor sizes and configurations to suit different sample volumes and types. The product is engineered to provide reliable and consistent performance in a laboratory setting.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Mab377, by Merck Group (2 mentions) Lsm 780, by Zeiss (2 mentions) Mabt868, by Merck Group (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Lsm 880, by Zeiss (2 mentions) Ab37615, by Abcam (1 mentions) Ab37612, by Abcam (1 mentions) A12379, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) T6793, by Merck Group (1 mentions) H3570, by Thermo Fisher Scientific (1 mentions) T6557, by Merck Group (1 mentions) Nb100 61071, by Novus Biologicals (1 mentions) Sc 240226, by Santa Cruz Biotechnology (1 mentions) Avicel rc 591, by FMC Biopolymer (1 mentions) Tpck treated trypsin, by Merck Group (1 mentions) A21428, by Thermo Fisher Scientific (1 mentions) Vectashield h 1000, by Vector Laboratories (1 mentions)

18 protocols using a 21311

Antibody Profiling of Centrosomal Proteins

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Commercial antibodies used were as follows: anti-Acetylated tubulin (^Actub, clone 6-11B-1, 1/10000, T6793, Sigma), anti-Gamma-tubulin (GTU-88, 1/1000, T6557, Sigma), anti-b-actin (clone AC-15, 1/30000, A5441, Sigma), anti-PACSIN1 (1/100, 196 003, Synaptic Systems), anti-PACSIN2 (1/250, ab37615, Abcam), anti-PACSIN2 (1/500, 10518-2-AP, Proteintech), anti-PACSIN3 (1/100, ab37612, Abcam), anti-Pericentrin (PCTN, 1/5000, NB100-61071, Novus Biologicals), anti-EHD1 (EPR4954, 1/500, ab109311, Novus Biologicals), anti-RPGRIP1L (1/200, 55160-1-AP, Proteintech), anti-TMEM67 (1/200, 12780-1-AP, Proteintech), anti-CEP164 (1/500, sc-240226, Santa Cruz), anti-CP110 (1/1000, 12780-1-AP, Proteintech), anti-CEP97 (1/1000, A301-945A, Bethyl), anti-Arl13b (1/300, clone N295B/66, NeuroMab/UC Davis), anti-GFP Alexa 488 (1/1000, A21311, Molecular Probes Life Technologies), Phalloidin conjugated with Alexa 488 (1/50, A12379, Molecular Probes Life Technologies), Hoechst (1/3000, H3570, Molecular Probes Life Technologies) and all secondary antibodies were from Life Technologies. The rabbit anti-RAB8A antibody was a gift from Johan Peränen (University Helsinki, Finland). SNAP-Cell647-SiR reagent was purchased from New England Biolabs. Nocodazole (Calbiochem) was from Sigma. Doxycyclin hydrochloride was obtained from Sigma and used according to manufacturer’s instruction.

Insinna C., Lu Q., Teixeira I., Harned A., Semler E.M., Stauffer J., Magidson V., Tiwari A., Kenworthy A.K., Narayan K, & Westlake C.J. (2019). Investigation of F-BAR domain PACSIN proteins uncovers membrane tubulation function in cilia assembly and transport. Nature Communications, 10, 428.

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Spatiotemporal Calcium Dynamics Imaging

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Variants were co-expressed with membrane-localized myr::tdTomato using the MB077B driver. Adults 3–6 days old were harvested, brains dissected and fixed using standard techniques. GCaMP variants were directly labelled with rabbit anti-GFP (1:500; AlexaFluor 488; A-21311, Molecular Probes). Primary rat anti-RFP (1:500; mAb 5F8, Chromotek) and secondary goat anti-rat Cy3 (1:1,000; 112-165-167, Jackson) labelled tdTomato.

Zhang Y., Rózsa M., Liang Y., Bushey D., Wei Z., Zheng J., Reep D., Broussard G.J., Tsang A., Tsegaye G., Narayan S., Obara C.J., Lim J.X., Patel R., Zhang R., Ahrens M.B., Turner G.C., Wang S.S., Korff W.L., Schreiter E.R., Svoboda K., Hasseman J.P., Kolb I, & Looger L.L. (2023). Fast and sensitive GCaMP calcium indicators for imaging neural populations. Nature, 615(7954), 884-891.

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Plaque Assay for Influenza A Virus Quantification

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MDCK cells were seeded at 5 x 10⁵ cells per well in a six-well plate. The next day, the cells were infected with a ten-fold dilution series of the virus in serum-free medium, in a total volume of 1 ml. The inoculum was removed after 1 h of incubation at 37°C and replaced by an overlay of 0.6% Avicel RC-591 (FMC Biopolymer) in serum-free medium containing 2 μg/ml TPCK-treated trypsin (Sigma). After three days of incubation at 37°C, the overlay was removed and the cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. The cells were stained with an anti-M2e mouse monoclonal antibody and the plaques were visualized with TrueBlue peroxidase substrate (KPL). For the determination of the percentage of GFP-positive cells, the plaques were first detected with an anti-GFP antibody (A21311; Molecular probes) and subsequently with a monoclonal anti-M2e antibody or a polyclonal anti-influenza A serum.

De Baets S., Verhelst J., Van den Hoecke S., Smet A., Schotsaert M., Job E.R., Roose K., Schepens B., Fiers W, & Saelens X. (2015). A GFP Expressing Influenza A Virus to Report In Vivo Tropism and Protection by a Matrix Protein 2 Ectodomain-Specific Monoclonal Antibody. PLoS ONE, 10(3), e0121491.

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Immunodetection of Myogenic Markers in Zebrafish Larvae

Cited 1 time

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Larvae were fixed with 2% paraformaldehyde in PBS for 25 min to overnight depending on the stage. Immunodetection for slow myosin heavy chain (MyHC) (1:5, F59; Devoto et al., 1996 (link)), general MyHC (1:10, A4.1025; Blagden et al., 1997 (link)), Pax7 (1:5; DSHB; Kawakami et al., 1997 (link)), Myogenin (1:50, sc-576, Santa Cruz) and GFP (1:500, TP-401, Torrey Pines or 1:500, G1546, Sigma) was performed in PBS with 0.5-1% Triton X-100 (PBT) for between overnight and 5 days at 4°C on a rotary shaker, depending on larval age and antigen (Hinits and Hughes, 2007 (link)) followed by Alexa Fluor 488 or 555 secondary antibodies (1:1000; A21121 and A21428, respectively; Invitrogen). at least overnight (±Hoechst 33342) at 4°C. After EdU detection in Fig. S4, samples were blocked in 5% BSA, 3% normal goat serum, PBT for 20 min, incubated using Alexa Fluor 488-conjugated anti-GFP (1:500, A-21311, Molecular Probes) and Hoechst in block buffer (shaking at 4°C for 3-6 h), followed by 6×15 min washes in PBT. Phalloidin-Alexa Fluor 555 or phalloidin-Alexa Fluor 633 (1:1000, A34055 or A22284, Thermo Fisher) were used to-stain F-actin. Larvae were mounted on glass slides under bridged coverslips in Citifluor AF1 or Vectashield (H-1000, Vector Laboratories). In situ mRNA hybridisation was as described (Groves et al., 2005 (link)).

Pipalia T.G., Koth J., Roy S.D., Hammond C.L., Kawakami K, & Hughes S.M. (2016). Cellular dynamics of regeneration reveals role of two distinct Pax7 stem cell populations in larval zebrafish muscle repair. Disease Models & Mechanisms, 9(6), 671-684.

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Immunocytochemistry of Patterned Neurons

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Neurons were replated on 50-μm grids patterned on the bottom of 35-mm MatTek dishes (MatTek CatP35G-0-14-C) previously coated with 100 μg/ml PLO and 5 μg/ml laminin. Immediately after live imaging, cells were fixed in situ with 4% paraformaldehyde (PFA, Core Bio Services) for 30 min at room temperature. The cells were then washed three times in phosphate buffered saline (PBS), permeabilized with 0.2% Triton X-100 in PBS for 15 min and blocked with 2% bovine serum albumin (BSA, Gemini Bio-products) in PBS for 2 h at room temperature. Primary antibodies (chicken anti-MAP2, Abcam ab5392, 1:500; rabbit anti-GFP, Molecular Probes A-21311, 1:1,000) were diluted in blocking buffer (2% BSA in PBS). Cells were incubated with the primary antibodies overnight at 4°C. The following day cells were washed three times in PBS, and incubated with the secondary antibodies coupled to Alexa Fluor 488, 555, and 647 (Life Technologies, 1:1,000 in blocking buffer) for 1 h at room temperature. Cells were then washed three times in PBS to remove the secondary antibodies, and DAPI (1:10,000 in PBS) was added for 30 min at room temperature to counterstain cell nuclei. Cells were then washed again three times in PBS. Finally, the PBS was aspirated, and the cells were submerged in a drop of ProLong Gold anti-fade mountant (Life Technologies) and covered with a glass coverslip for subsequent analysis.

Puppo F., Sadegh S., Trujillo C.A., Thunemann M., Campbell E.P., Vandenberghe M., Shan X., Akkouh I.A., Miller E.W., Bloodgood B.L., Silva G.A., Dale A.M., Einevoll G.T., Djurovic S., Andreassen O.A., Muotri A.R, & Devor A. (2021). All-Optical Electrophysiology in hiPSC-Derived Neurons With Synthetic Voltage Sensors. Frontiers in Cellular Neuroscience, 15, 671549.

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Immunohistochemical Analysis of Brain Sections

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Brain sections were stained with anti-GFP (A-21311, Life Technologies) and anti-CD31 (MEC 13.3, BD Pharmingen) as described (20 (link)). Sections were counterstained in DAPI (Sigma-Aldrich) then cover-slipped in ProLong Diamond anti-fade Mountant (Life Technologies).

Shaw T.N., Inkson C.A., Villegas-Mendez A., Pattinson D.J., Strangward P., Else K.J., Draper S.J., Zeef L.A, & Couper K.N. (2019). Infection-Induced Resistance to Experimental Cerebral Malaria Is Dependent Upon Secreted Antibody-Mediated Inhibition of Pathogenic CD8+ T Cell Responses. Frontiers in Immunology, 10, 248.

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Marmoset Brain Immunofluorescence Imaging

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Marmosets were deeply anesthetized and perfused intracardially with 4% paraformaldehyde in phosphate buffer. The brain was post-fixed and then cryoprotected with 30% sucrose at 4°C. Coronal sections of 40-µm-thickness encompassing the regions of interest were prepared from each hemisphere. For immunofluorescence, free-floating sections were blocked with blocking solution (10% normal donkey serum and 0.3% Triton X-100 in PBS, pH 7.4) for 1 h at room temperature, followed by overnight incubation with anti-neuronal nuclear protein (NeuN) antibody (1:1000; MAB377, EMD Millipore) and glial fibrillary acidic protein (GFAP) anti-body (1:1000; G3893, Sigma-Aldrich). The primary antibody was detected using donkey anti-mouse secondary antibody conjugated with Alexa Fluor^® 546 (1:1000; A10036, Life Technologies) in combination with rabbit anti-green fluorescence protein (GFP) antibody conjugated with Alexa Fluor^® 488 (1:1000; A21311, Life Technologies) for 2 h at room temperature. Nuclei were counterstained with 4′,6-Diamidino-2-phenylindole (DAPI; D9542, Sigma-Aldrich). Sections were visualized using a confocal microscope (LSM 780, Carl Zeiss).

Santisakultarm T.P., Kersbergen C.J., Bandy D.K., Ide D.C., Choi S.H, & Silva A.C. (2016). Two-photon imaging of cerebral hemodynamics and neural activity in awake and anesthetized marmosets. Journal of neuroscience methods, 271, 55-64.

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Immunolabeling of EGFP-Expressing Cells

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Two primary antibodies were used to detect EGFP either to intensify the native fluorescent signal or to visualize the expression by diaminobenzadine (DAB) staining. These antibodies are listed in Table 2. Secondary antibodies are listed in Table 3. Two anti-GFP antibodies were used in this study. The first, A-21311 (Life Technologies, RRID:AB_10058149), is a rabbit polyclonal antibody raised against Aequorea victoria GFP, and is conjugated to Alexa Fluor 488. In this study, however, this antibody was used only for DAB staining by way of a biotin-conjugated secondary antibody. The second anti-GFP antibody used in this study, (Life Technologies Cat#A-11122, RRID:AB_10073917), is also a rabbit polyclonal antibody raised against A. victoria GFP, and was used for immunofluorescence by way of fluorophore-conjugated secondary antibodies. No differences in the pattern of expression were noted between the two anti-GFP antibodies used in this study. Neither anti-GFP antibody used in the study reacted with tissue from wild type littermates (data not shown). One antibody, NeuN (Millipore Cat#MAB377B, RRID:AB_177621), was used to identify neuroanatomical structures in the brain. This antibody has been used extensively as an anatomical marker of most neuronal cell types, with 124 citations listed in the most recent Journal of Comparative Neurology Antibody Database (V14; RRID:nlx_143660).

Condro M.C., Matynia A., Foster N.N., Ago Y., Rajbhandari A.K., Van C., Jayaram B., Parikh S., Diep A.L., Nguyen E., May V., Dong H.W, & Waschek J.A. (2016). High-resolution characterization of a PACAP-EGFP transgenic mouse model for mapping PACAP-expressing neurons. The Journal of comparative neurology, 524(18), 3827-3848.

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Immunofluorescence Staining of Myogenic Markers

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The following primary and secondary antibodies were used to stain cells and tissue sections for immunofluorescence in this study: Pax7 (1:50; Developmental Studies Hybridoma Bank), MyoD (1:200; C-20, Santa Cruz Biotechnology), myosin heavy chain (1:200; M4276, Sigma-Aldrich), desmin (1:200; H-76, Santa Cruz Biotechnology), Alexa Fluor 488–conjugated anti-GFP (1:250; A-21311, Thermo Fisher Scientific), Alexa Fluor 488 goat anti-mouse immunoglobulin G (IgG; 1:250; A-11029, Thermo Fisher Scientific), Alexa Fluor 488 goat anti-rabbit IgG (1:250; A-11034, Thermo Fisher Scientific), Alexa Fluor 555 goat anti-mouse IgG (1:250; A-21422, Thermo Fisher Scientific), and Alexa Fluor 555 goat anti-rabbit IgG (1:250; A-21424, Thermo Fisher Scientific). Alexa Fluor 555 phalloidin (1:50; Thermo Fisher Scientific) and Alexa Fluor 647 phalloidin (1:50; Thermo Fisher Scientific) were used to stain F-actin. Hoechst 33342 (1:1000; Thermo Fisher Scientific) and DRAQ5 (1:1000; Thermo Fisher Scientific) were used to counterstain cell nuclei.

Han W.M., Anderson S.E., Mohiuddin M., Barros D., Nakhai S.A., Shin E., Amaral I.F., Pêgo A.P., García A.J, & Jang Y.C. (2018). Synthetic matrix enhances transplanted satellite cell engraftment in dystrophic and aged skeletal muscle with comorbid trauma. Science Advances, 4(8), eaar4008.

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Whole-mount Immunostaining of Embryos

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Euthanized embryos were fixed in a solution containing 4% paraformaldehyde in PBS for at least 2 h at room temperature. Following fixation, samples were washed with 0.3% PBSTx (

3 \times 5

min), permeabilized with acetone (100% acetone, 10 min incubation at

- 20^{\circ}

C), washed with 0.3% PBSTx (

3 \times 10

min) and blocked in 0.1% BSA/0.3% PBSTx for 2 h. Embryos were incubated with acetylated tubulin antibody (clone 6-11B-1, MABT868, Sigma-Aldrich, 1:1000) overnight at

4^{\circ}

C. On the next day samples were washed (0.3% PBSTx,

3 \times 1

h) and incubated with the secondary antibody (Alexa-labeled goat-anti-mouse 555 plus, A32727, Thermo Fisher Scientific, 1:1000), an anti-GFP antibody conjugated with Alexa 488 (A-21311, Thermo Fisher Scientific, 1:1000) and DAPI (D1306, Thermo Fisher Scientific, 1:1000) overnight at

4^{\circ}

C. The next day samples were washed (0.3% PBSTx,

3 \times 1

h) and transferred to a series of increasing glycerol concentrations (25%, 50%, and 75%).

Hansen J.N., Rassmann S., Stüven B., Jurisch-Yaksi N, & Wachten D. (2021). CiliaQ: a simple, open-source software for automated quantification of ciliary morphology and fluorescence in 2D, 3D, and 4D images. The European Physical Journal. E, Soft Matter, 44(2), 18.

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