A21131

Manufactured by Thermo Fisher Scientific

Sourced in France, United States

The A21131 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is a multi-function device designed for various applications in scientific research and analysis. The core function of the A21131 is to perform accurate measurements and data processing tasks. Further details about its intended use or specific applications are not available.

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Lab products found in correlation

Lsm 710 confocal microscope, by Zeiss (6 mentions) A2547, by Merck Group (3 mentions) A 21124, by Thermo Fisher Scientific (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Ab71916, by Abcam (2 mentions) Ab28364, by Abcam (2 mentions) Anti rabbit igg af546, by Thermo Fisher Scientific (2 mentions) Dmi8 thunder imager fluorescent microscope, by Leica (2 mentions) Lsm 780, by Zeiss (2 mentions) Ab76956, by Abcam (1 mentions) Ab52917, by Abcam (1 mentions) Ab15191, by Abcam (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) A11012, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 conjugated goat polyclonal anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Mab1195, by R&D Systems (1 mentions) A12381, by Thermo Fisher Scientific (1 mentions) 1x71 microscope, by Olympus (1 mentions) A21123, by Thermo Fisher Scientific (1 mentions)

10 protocols using a21131

Multiparametric Immunofluorescence Assay

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2D cultures or MBs were washed in PBS and incubated with a 5 μM NucView 488 caspase-3 substrate (Interchim, Montluçon, France) diluted in PBS. After washing with PBS, HMSCs were fixed with a 4% (w/v) PFA (Alpha Aesar, Heysham, UK) for 30 min and permeabilized with 0.2 to 0.5% (v/v) Triton X-100 (Sigma-Aldrich) for 5 min. The samples were blocked with 5% (v/v) FBS in PBS for 30 min and incubated with a rabbit polyclonal anti–COX-2 primary antibody (ab15191, Abcam, Cambridge, UK) diluted at 1:100 in 1% (v/v) FBS for 4 hours. After washing with PBS, the samples were incubated with an Alexa Fluor 594–conjugated goat polyclonal anti-rabbit IgG secondary antibody (A-11012, Life Technologies, Saint Aubin, France) diluted at 1:100 in 1% (v/v) FBS for 90 min. Last, the cells were counterstained with 0.2 μM DAPI for 5 min (Sigma-Aldrich) and then washed with PBS.
The same protocol was used for the staining of VEGF-A–expressing cells using a rabbit anti-human VEGF-A monoclonal antibody (ab52917, Abcam, Cambridge, UK), which was revealed using the same secondary antibody as above. RUNX-2–positive cells were similarly stained using a mouse anti-human RUNX-2 monoclonal antibody (ab76956, Abcam, Cambridge, UK), which was revealed using an Alexa Fluor 488 goat anti-mouse IgG2a secondary antibody (A-21131, Life Technologies, Saint Aubin, France), both diluted at 1:100 in 1% (v/v) FBS.

Sart S., Tomasi R.F., Barizien A., Amselem G., Cumano A, & Baroud C.N. (2020). Mapping the structure and biological functions within mesenchymal bodies using microfluidics. Science Advances, 6(10), eaaw7853.

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Quantifying Nerve Innervation in Tissue Sections

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Tissue was harvested and fixed in 4% paraformaldehyde in PBS, brought through a sucrose series (beginning with incubation in 5 % sucrose and ending with a final 30 % sucrose equilibration step), embedded in OCT, and flash-frozen. Frozen sections were acquired via cryostat at 16 μm. Immunohistochemistry with the mouse monoclonal (IgG_2a) β-III tubulin antibody (1:50 of 0.5 mg/mL stock, R&D Systems: MAB1195, LOT: HGQ0113121, Clone # TuJ-1) was performed as in (30 (link)). Goat anti-mouse IgG_2a coupled to Alexa-fluor 488 (1:300, Life Technologies: A21131), and phalloidin conjugated to Alexa-fluor 594 (1:40, Life Technologies: A12381) were used secondarily. Samples were incubated in DAPI (1.4 μmol/L) for 10 minutes, and slides were mounted with hydromount. Fluorescent images were acquired on an Olympus 1X71 microscope and processed with MetaMorph Basic (V7.7.0.0) software, and low magnification images were manually stitched together. ImageJ was used to add scale bars, adjust color balance, and quantify images. For quantification of innervation in Figure 3C, a common threshold for fluorescence emitted by nerves was applied to all images using ImageJ. The “Analyze Particles” function within ImageJ was used to measure surface area covered by nerves. Total cross-sectional area of sections was also measured with ImageJ as with muscle and bone.

Bryant D.M., Sousounis K., Farkas J.E., Bryant S., Thao N., Guzikowski A.R., Monaghan J.R., Levin M, & Whited J.L. (2017). Repeated removal of developing limb buds permanently reduces appendage size in the highly-regenerative axolotl. Developmental biology, 424(1), 1-9.

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Immunofluorescent Staining and Laser Microdissection

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Cut sections were fixed in chilled 4% PFA for 3 mins before permeablisation in a methanol series (70% MeOH 10 min, 95% MeOH 10 min, 100% MeOH 20 min, 95% MeOH 10 min, 70% MeOH 10 min). Sections were washed in TTBS and blocked in 10% NGS TTBS for 1 hour at room temperature (or 10% donkey serum (DS) for anti-nanog staining). 1° antibodies were against nestin (1:1000, Abcam, cat# ab6320), alpha fetoprotein (1:100, Abcam, cat# ab133617), smooth muscle actin (1:100, Abcam, cat# ab125044), nanog (1:40, R&D systems, cat# AF1997SP), COXIV + COXIVL2 (1:500, Abcam, cat# ab110261). Antibodies were diluted in 3% NGS TTBS and incubated overnight at 4 °C. Anti-nanog was diluted in 3% DS TTBS. Sections were washed 3× in TTBS and appropriate 2° applied in 3% NGS or DS TTBS with 10 µg/ml Hoechst for 90 min at room temperature. 2° antibodies used were all purchased from Thermo Fisher (cat#; A11056, A21123, A21131, A11010). After staining slides were washed 3× in TTBS and either mounted with ProLong Gold mountant on coverslips for imaging or washed 3× in ultrapure water and allowed to air dry in the dark prior to microdissection. Germlayers were laser microdissected using a Zeiss PALM microbeam. Dissected regions were lysed in 20 µl single cell lysis buffer and lysed as described above.

Russell O.M., Fruh I., Rai P.K., Marcellin D., Doll T., Reeve A., Germain M., Bastien J., Rygiel K.A., Cerino R., Sailer A.W., Lako M., Taylor R.W., Mueller M., Lightowlers R.N., Turnbull D.M, & Helliwell S.B. (2018). Preferential amplification of a human mitochondrial DNA deletion in vitro and in vivo. Scientific Reports, 8, 1799.

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Fluorescent Imaging of Lung Cell Populations

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Formalin-fixed and paraffin-embedded lung sections from mock- and IAV-infected mice were prepared as previously described (52 (link)) and probed for epithelial or endothelial cell detection using monoclonal mouse anti–α-smooth muscle actin (αSMA) (1:10,000; A2547, Sigma-Aldrich, Darmstadt, Germany), rabbit anti-mouse CD31 (1:50; ab28364, Abcam, Cambridge, UK), or rabbit anti-mouse EpCAM (1:50; ab71916, Abcam, Cambridge, UK) antibodies. Tissues were incubated with corresponding secondary antibodies goat anti-mouse immunoglobulin G2a (IgG2a)–AF488 (1:200; A-21131, Thermo Fisher Scientific) and anti-rabbit IgG-AF546 (1:200; Invitrogen, Carlsbad, CA) in 7% goat serum/PBS for 1 hour at room temperature, and nuclei were counterstained with 4′,6-diamidino-2-phenylindole. All the images were acquired using a Leica DMi8 Thunder Imager fluorescent microscope and analyzed using the Fiji/ImageJ software. For colocalization analysis, a plot profile tool was used to measure and generate the intensity plots of the boxed regions, and Coloc2 plug-in was used to quantify colocalization parameters using Pearson’s correlation efficiency recorded in Fig. 7 and fig. S8.

Guo K., Yombo D.J., Wang Z., Navaeiseddighi Z., Xu J., Schmit T., Ahamad N., Tripathi J., De Kumar B., Mathur R., Hur J., Sun J., Olszewski M.A, & Khan N. (2024). The chemokine receptor CXCR3 promotes CD8+ T cell–dependent lung pathology during influenza pathogenesis. Science Advances, 10(1), eadj1120.

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Visualizing Lung Cell Populations in IAV Infection

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Formalin-fixed and paraffin-embedded lung sections from mock and IAV-infected mice were prepared as previously described (47) and probed for epithelial or endothelial cells detection using monoclonal mouse anti-αSMA (1:10,000; A2547; Sigma-Aldrich, Darmstadt, Germany), rabbit anti-mouse CD31 (1:50; ab28364, Abcam, Cambridge, UK), or rabbit anti-mouse EpCAM (1:50, ab71916, Abcam, Cambridge, UK) antibodies. Tissues were incubated with corresponding secondary antibodies goat anti-mouse IgG2a-AF488 (1:200; A-21131, ThermoFisher), anti-rabbit IgG-AF546 (1:200; Invitrogen, Carlsbad, CA), in 7% goat serum/PBS for 1h at RT, nuclei were counterstained with DAPI. All the images were acquired using a Leica DMi8 Thunder Imager fluorescent microscope and analyzed using Image J software.

Guo K., Yombo D.J.K., Xu J., Wang Z., Schmit T., Tripathi J.K., Hur J., Sun J., Olszewski M.A., & Khan M.N. (2022). The chemokine receptor CXCR3 promotes CD8⁺T cell-dependent lung pathology during influenza pathogenesis.

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Immunofluorescence Analysis of RABV-Infected Mouse Brain

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Brain tissues harvested from rRABV-infected mice at 5 dpi were fixed in 10% phosphate-buffered formalin for more than 48 h and then embedded into paraffin blocks. The paraffin blocks were sectioned at a 4-μm thickness and mounted on Platinum PRO micro glass slides (Matsunami). The sections on slides were subjected to antigen retrieval in citrate buffer for 5 min by a pressure cooker. The slides were then treated with 10% goat serum (Nichirei Biosciences) for 1 h at room temperature and incubated at 4 °C overnight with primary antibodies against the following proteins: NeuN (ab104224; Abcam, 1:1000), GFAP (G9269; Sigma Aldrich, 1:400) or RABV N protein (3R7-5B12; Hytest, 1:100). After three-times washes with PBST (0.01% Tween 20 in PBS), secondary staining was performed with Alexa Fluor 488-anti-mouse IgG2a antibody (A-21131; Invitrogen, 1:1000), Alexa Fluor Plus 594-anti-rabbit IgG (H+L) antibody (A32740; Invitrogen, 1:1000) and Alexa Fluor 647-anti-mouse IgG2b antibody (A-21242; Invitrogen, 1:1000) in the presence of 10 μg/mL Hoechst 33,342 for 1 h at room temperature. Microscopic analysis was conducted by LSM780 and ZEN software (Zeiss).

Itakura Y., Tabata K., Morimoto K., Ito N., Chambaro H.M., Eguchi R., Otsuguro K.I., Hall W.W., Orba Y., Sawa H, & Sasaki M. (2022). Glu333 in rabies virus glycoprotein is involved in virus attenuation through astrocyte infection and interferon responses. iScience, 25(4), 104122.

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Immunofluorescence Analysis of Cell Markers

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Upon completion of the stretching experiments, the samples were fixed with 3.7% formaldehyde for 20 min at room temperature, and permeabilized with 0.5% Triton X-100 in PBS for 3 min. Non-specific antibody binding was blocked 30 min with 3% BSA in PBS. The samples were subsequently incubated with primary antibodies in 1.5% BSA in PBST at 4°C overnight. The primary antibodies used in this study were alpha smooth muscle actin (αSMA) (A2547, Sigma-Aldrich, 1:600), KI67 (PA5-16446, Invitrogen, 1:100) and collagen I (C2456, Sigma-Aldrich, 1:100). The samples were washed three times for 5 min with PBST and incubated with secondary antibodies in PBST for 1 h at room temperature. The secondary antibodies used were goat anti-mouse IgG2a Alexa Fluor 488 (A21131, Invitrogen, 1:1,000), goat anti-rabbit IgG Alexa Fluor 555 (A21428, Invitrogen, 1:500) and goat anti-mouse IgG1 Alexa Fluor 488 (A21121, Invitrogen, 1:500). All samples were stained for DAPI (Sigma-Aldrich, 1:500) to visualize cell nuclei. Subsequently, the samples were mounted with Mowiol (Sigma-Aldrich) on thin coverslips. Image acquisition was done using an inverted fluorescence microscope (Leica DMi8) with ×20 magnification. Negative controls of immunofluorescence (IF) staining are presented in Supplementary Figure S1.

Karakaya C., van Turnhout M.C., Visser V.L., Ristori T., Bouten C.V., Sahlgren C.M, & Loerakker S. (2022). Notch signaling regulates strain-mediated phenotypic switching of vascular smooth muscle cells. Frontiers in Cell and Developmental Biology, 10, 910503.

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Immunostaining of Cochlear and Neural Samples

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The cochleae were dissected in cold Hanks Balanced Salt Solution (HBSS) in order to prevent protein degradation and then fixed with 4% paraformaldehyde (PFA) for 1 h at room temperature. In vitro cultured neurospheres were also fixed with 4% PFA for 1 h at room temperature. After washing with phosphate buffered saline with tween (PBST) three times, the cochleae or neurospheres were blocked with blocking solution for 1 h at room temperature and then incubated overnight at 4°C with primary antibodies. The primary antibodies used were anti-Myosin7a (Myo7a; Proteus Bioscience, #25-6790; 1:1,000 dilution), anti-Sox2 (1:400 dilution), anti-AFF1 (1:400 dilution), anti-AFF4 (1:50 dilution), and anti-ELL3 (1:400 dilution). After washing again three times, the cochleae or neurospheres were further incubated with secondary antibodies (Invitrogen, A21131, A21124) diluted 1:400 in PBT2 for 1 h at room temperature. After washing three times, the cochleae or neurospheres were mounted on slides with anti-fade fluorescence mounting medium (DAKO, S3023). Images were captured by Zeiss LSM 710 confocal microscope and analyzed by Image J software.

Chen Y., Qiang R., Zhang Y., Cao W., Wu L., Jiang P., Ai J., Ma X., Dong Y., Gao X., Li H., Lu L., Zhang S, & Chai R. (2021). The Expression and Roles of the Super Elongation Complex in Mouse Cochlear Lgr5+ Progenitor Cells. Frontiers in Cellular Neuroscience, 15, 735723.

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Immunostaining of Cochlear and Neural Cells

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The cochleae were dissected in cold HBSS in order to prevent protein degradation and then xed with 4% PFA for 1 h at room temperature. In vitro cultured neurospheres were also xed with 4% PFA for 1 h at room temperature. After washing with PBST three times, the cochleae or neurospheres were blocked with blocking solution for 1 h at room temperature and then incubated overnight at 4°C with primary antibodies. The primary antibodies used were anti-Myosin7a (Myo7a; Proteus Bioscience, #25-6790; 1:1000 dilution), anti-Sox2 (1:400 dilution), anti-AFF1 (1:400 dilution), anti-AFF4 (1:50 dilution), and anti-ELL3 (1:400 dilution). After washing again three times, the cochleae or neurospheres were further incubated with secondary antibodies (Invitrogen, A21131, A21124) diluted 1:400 in PBT2 for 1 h at room temperature. After washing three times, the cochleae or neurospheres were mounted on slides with antifade uorescence mounting medium (DAKO, S3023). Images were captured by Zeiss LSM 710 confocal microscope and analyzed by Image J software.

Chen Y., Qiang R., Zhang Y., Cao W., Wu L., Jiang P., Ai J., Ma X., Dong Y., Gao X., Li H., Lü L., Zhang S., & Chai R. (2021). The Expression and Roles of the Super Elongation Complex in Mouse Cochlear Lgr5+ Progenitor Cells.

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Quantification of Synaptic Ribbons in Cochlear Tissues

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After the final physiological tests, the animals were sacrificed, and their cochleae were used for quantification of the synaptic ribbons, according to our previously published protocols (Liu et ). Briefly, after fixing with 4% paraformaldehyde in phosphate-buffered saline (PBS), the cochlear tissues were dissected, permeabilized with 1% Triton X-100 in PBS for 1 hour, incubated in 5% goat serum in PBS for a further 1 hour, and then incubated overnight at 4 °C with primary antibodies against both C-terminal binding protein 2 (CtBP2) and post-synaptic density-95 (PSD-95) (mouse IgG1 to CtBP2; BD Biosciences, Franklin Lakes, NJ, USA: cat. # 612044, 1:200; mouse IgG2a to PSD95; Millipore, Billerica, MA, USA: cat. # MAB1596, 1:600). After the reaction, the tissues were washed and treated with the corresponding secondary antibodies (A21124 and A21131, respectively; Invitrogen, Carlsbad, CA, USA) for 2 h at room temperature, and then mounted on microscope slides. ) using a confocal laser-scanning microscope (LSM 710 META; Zeiss, Shanghai, China) with a 63× water-immersion objective. Image stacks were then exported to ImageJ image-processing software (National Institutes of Health, Bethesda, MD, USA), and over 10 successive IHCs at each frequency position of the cochlea were selected to count the puncta of CtBP2 (red) and PSD95 (green).

Chen H., Xing Y., Zhang Z., Tao S., Wang H., Aiken S., Yin S., Yu D, & Wang J. (2019). Coding-in-Noise Deficits are Not Seen in Responses to Amplitude Modulation in Subjects with cochlear Synaptopathy Induced by a Single Noise Exposure. Neuroscience, 400.

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