Incubation:Article Title: Activin A promotes bone fracture repair and acts through a novel myofibroblastic cell population in callus
Article Snippet: .. Sections were incubated with primary antibodies, such as mouse anti-αSMA, goat anti-Activin A (R&D, AF338), at 4°C overnight, followed by Alexa Fluor 488 donkey anti-mouse, Alexa Fluor 647 donkey anti-goat (Invitrogen, A-21447) secondary antibodies incubation 1 hour at RT. ..
Article Title: Acute Intermittent Hypoxia Exerts Beneficial Effects and Promotes Repair in Male Mice in the Experimental Autoimmune Encephalomyelitis Model of Multiple Sclerosis.
Article Snippet: Slides were washed in 0.1 M PBS and blocked with SeaBlock (Thermo Scientific; Rockford, IL, USA; cat. #37527) for 1 h at room temperature. .. Slides were then incubated with primary antibodies (diluted in 0.1% Triton- X in 0.1 M PBS, 10% SeaBlock [overnight at 4°C]): goat anti- activin- A (R&D Systems #AF338; 1:25), rabbit anti- βIII- Tubulin (Sigma- Aldrich #T2200; 1:500), rat anti- ED- 1/CD68 (Abcam #ab53444; 1:500), rabbit anti- ED- 1/ CD68 (Abcam #ab125212; 1:600), rabbit anti- inducible nitric oxide synthase (iNOS; Sigma- Aldrich #N7782; 1:500), rat antimyelin basic protein (MBP; NovusBio #NB600- 717; 1:20), goat anti- platelet derived growth factor receptor alpha (PDGFRα; R&D Systems #AF1062; 1:75), mouse anti- SMI31 (Cedarlane #SMI- 31R; 1:1000). .. After rinsing with 0.1 M PBS, secondary antibodies (diluted in 0.01% Triton- X in 0.1 M PBS, 1% SeaBlock [1 h room temperature]) were applied: goat anti- mouse Alexa488 (Abcam #150117; 1:1000), mouse anti- rat Alexa488 (Jackson ImmunoResearch #212- 545- 106; 1:1000), donkey anti- rabbit CY3 (Jackson ImmunoResearch #711- 166- 152; 1:3000), donkey anti- goat DyLight488 (Jackson ImmunoResearch #705- 485- 003; 1:1000).
Article Title: Activin A marks a novel progenitor cell population during fracture healing and reveals a therapeutic strategy
Article Snippet: .. Sections were incubated with mouse anti-αSMA, goat anti-Activin A (R&D, AF338) primary antibodies at 4°C overnight, followed by incubation with Alexa Fluor 488 donkey anti-mouse (Invitrogen, A-21202) and Alexa Fluor 647 donkey anti-goat (Invitrogen, A-21447) secondary antibodies for 1 hr at RT. .. Fluorescence images were captured by a Zeiss LSM 710 scanning confocal microscope interfaced with the Zen 2012 software (Carl Zeiss Microimaging LLC).
Article Title: Activin A marks a novel progenitor cell population during fracture healing and reveals a therapeutic strategy
Article Snippet: .. Sections were incubated with mouse anti-αSMA, goat anti- Activin A (R&D, AF338) primary antibodies at 4°C overnight, followed by incubation with Alexa Fluor 488 donkey anti- mouse (Invitrogen, A- 21202) and Alexa Fluor 647 donkey anti- goat (Invitrogen, A- 21447) secondary antibodies for 1 hr at RT. .. Fluorescence images were captured by a Zeiss LSM 710 scanning confocal microscope interfaced with the Zen 2012 software (Carl Zeiss Microimaging LLC).
other:Article Title: Activity-dependent regulation of microglia numbers by pyramidal cells during development shape cortical functions
Article Snippet: The following primary antibodies were used: goat anti-Activin A (1:100, R&D Systems, catalog no. AF338), mouse anti-Bassoon (1:500, Abcam, catalog no. AB82958), rabbit anti-Homer (1:500, Synaptic Systems, catalog no. 160 002), chicken anti-GFP (green fluorescent protein) (1:3000, Aves Lab, catalog no. GFP-1020), goat anti-Iba1 (1:500, Biotechne, catalog no. NB100-1028), guinea pig anti-Iba1 (1:500, Synaptic Systems, catalog no. 234 004), rat anti-RFP (red fluorescent protein) (1:500, Chromotek, catalog no. 5f8-100), rabbit anti-Ki67 (1:500, Abcam, catalog no. ab16667), rat anti-P2RY12 (1:100, BioLegend, catalog no. 848002), rabbit anti-pSMAD3 (1:200, Abcam, catalog no. ab52903), goat anti-Sox9 (1:200, Biotechne, catalog no. AF3075), and rabbit anti-Acvr1b (1:100, Thermo Fisher Scientific, catalog no. Ma5-38039).
Article Title: Activity-dependent regulation of microglia numbers by pyramidal cells during development shape cortical functions.
Article Snippet: The following primary antibodies were used: goat anti- Activin A (1:100, R&D Systems, catalog no. AF338), mouse anti-Bassoon (1:500, Abcam, catalog no. AB82958), rabbit anti- Homer (1:500, Synaptic Systems, catalog no. 160 002), chicken anti- GFP (green fluorescent protein) (1:3000, Aves Lab, catalog no. GFP- 1020), goat anti- Iba1 (1:500, Biotechne, catalog no. NB100- 1028), guinea pig anti- Iba1 (1:500, Synaptic Systems, catalog no. 234 004), rat anti- RFP (red fluorescent protein) (1:500, Chromotek, catalog no. 5f8- 100), rabbit antiKi67 (1:500, Abcam, catalog no. ab16667), rat anti- P2RY12 (1:100, BioLegend, catalog no. 848002), rabbit anti- pSMAD3 (1:200, Abcam, D ow nloaded from https://w w w .science.org on February 25, 2025 Kumaraguru et al., Sci.
Derivative Assay:Article Title: Acute Intermittent Hypoxia Exerts Beneficial Effects and Promotes Repair in Male Mice in the Experimental Autoimmune Encephalomyelitis Model of Multiple Sclerosis.
Article Snippet: Slides were washed in 0.1 M PBS and blocked with SeaBlock (Thermo Scientific; Rockford, IL, USA; cat. #37527) for 1 h at room temperature. .. Slides were then incubated with primary antibodies (diluted in 0.1% Triton- X in 0.1 M PBS, 10% SeaBlock [overnight at 4°C]): goat anti- activin- A (R&D Systems #AF338; 1:25), rabbit anti- βIII- Tubulin (Sigma- Aldrich #T2200; 1:500), rat anti- ED- 1/CD68 (Abcam #ab53444; 1:500), rabbit anti- ED- 1/ CD68 (Abcam #ab125212; 1:600), rabbit anti- inducible nitric oxide synthase (iNOS; Sigma- Aldrich #N7782; 1:500), rat antimyelin basic protein (MBP; NovusBio #NB600- 717; 1:20), goat anti- platelet derived growth factor receptor alpha (PDGFRα; R&D Systems #AF1062; 1:75), mouse anti- SMI31 (Cedarlane #SMI- 31R; 1:1000). .. After rinsing with 0.1 M PBS, secondary antibodies (diluted in 0.01% Triton- X in 0.1 M PBS, 1% SeaBlock [1 h room temperature]) were applied: goat anti- mouse Alexa488 (Abcam #150117; 1:1000), mouse anti- rat Alexa488 (Jackson ImmunoResearch #212- 545- 106; 1:1000), donkey anti- rabbit CY3 (Jackson ImmunoResearch #711- 166- 152; 1:3000), donkey anti- goat DyLight488 (Jackson ImmunoResearch #705- 485- 003; 1:1000).
Staining:Article Title: Injury of Adult Zebrafish Expressing Acvr1l Q204D Does Not Result in Heterotopic Ossification
Article Snippet: .. For Safranin O staining, slides were submerged in 0.02% Fast Green in 0.2% acetic acid for 15 s, followed directly by submersion in 1% Safranin O for 30 s. IHC analyses were performed as previously described 26 using the following antibodies: 1:200 goat anti-activin A (AF338; R&D Systems); 1:100 goat anti-Collagen I (AB758; Millipore); 1:200 rabbit anti-Collagen II (ab34712; Abcam, Cambridge, MA); 1:250 rabbit anti-pSmad2 (AB3849-I; Millipore); 1:50 rabbit anti-von Willebrand factor (vWF) (HPA001815; Sigma-Aldrich, St. Louis, MO); 1:500 donkey anti-rabbit and 1:500 donkey anti-goat (Jackson ImmunoResearch Laboratories, West Grove, PA). .. Stained sections were analyzed and imaged using a Zeiss AxioPhot epifluorescence microscope (Zeiss) and Axiocam 503 color camera (Zeiss) and processed using AxioVision SE64 microscopy software (Zeiss).
Immunohistochemistry:Article Title: Injury of Adult Zebrafish Expressing Acvr1l Q204D Does Not Result in Heterotopic Ossification
Article Snippet: .. For Safranin O staining, slides were submerged in 0.02% Fast Green in 0.2% acetic acid for 15 s, followed directly by submersion in 1% Safranin O for 30 s. IHC analyses were performed as previously described 26 using the following antibodies: 1:200 goat anti-activin A (AF338; R&D Systems); 1:100 goat anti-Collagen I (AB758; Millipore); 1:200 rabbit anti-Collagen II (ab34712; Abcam, Cambridge, MA); 1:250 rabbit anti-pSmad2 (AB3849-I; Millipore); 1:50 rabbit anti-von Willebrand factor (vWF) (HPA001815; Sigma-Aldrich, St. Louis, MO); 1:500 donkey anti-rabbit and 1:500 donkey anti-goat (Jackson ImmunoResearch Laboratories, West Grove, PA). .. Stained sections were analyzed and imaged using a Zeiss AxioPhot epifluorescence microscope (Zeiss) and Axiocam 503 color camera (Zeiss) and processed using AxioVision SE64 microscopy software (Zeiss).
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