Retinas were fixed and counterstained with the following antibodies. Primary antibodies: Goat anti-ChAT (Choline acetyltransferase; 1:200, Millipore Sigma #AB144); Rabbit anti-VGluT3 (1:250, Invitrogen #PA5-85784). Chicken anti-GFP (1:1000, Abcam #ab13970) was used to enhance the fluorescence of the Cre-dependent GFP virus. Rabbit anti-HA tag (1: 200, Cell Signaling Technology #3724) was used to stain the HA-tagged hM4Di receptor in the VGluT3 x DREADD mouse. Secondary antibodies: Donkey anti-Chicken 488 (1:1000, Jackson Immunoresearch #703-545-155); Donkey anti-Chicken 594 (1:1000, Jackson Immunoresearch #703-585-155); Donkey anti-Goat 488 (1:200, Invitrogen #A-11055); Donkey anti-Goat 594 (1:200, Invitrogen #A-11058); Donkey anti-Rabbit 647 (1:200, Invitrogen #A31573).
Donkey anti goat 594
Manufactured by Thermo Fisher Scientific
Sourced in United States
Donkey anti-goat 594 is a secondary antibody used in immunofluorescence applications. It is raised in donkeys and binds to goat primary antibodies, allowing detection of target proteins through fluorescent labeling.
Automatically generated - may contain errors
Lab products found in correlation
Donkey anti goat 488, by Thermo Fisher Scientific (2 mentions)
Donkey anti rabbit 488, by Thermo Fisher Scientific (2 mentions)
Donkey anti rabbit 647, by Thermo Fisher Scientific (2 mentions)
Rabbit anti ha tag, by Cell Signaling Technology (1 mentions)
Donkey anti chicken 488, by Jackson ImmunoResearch (1 mentions)
Claudin 5, by Thermo Fisher Scientific (1 mentions)
Donkey anti rabbit 546, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
Ab77095, by Abcam (1 mentions)
Ab97959, by Abcam (1 mentions)
Sc 8628, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse 594, by Thermo Fisher Scientific (1 mentions)
Mab4301, by Merck Group (1 mentions)
Mab4381, by Merck Group (1 mentions)
Fish skin gelatin, by Merck Group (1 mentions)
Rabbit anti tmem119, by Abcam (1 mentions)
Donkey anti rabbit 555, by Thermo Fisher Scientific (1 mentions)
Dapi counterstain, by Southern Biotech (1 mentions)
Goat anti epha4, by R&D Systems (1 mentions)
Rabbit anti ccr2, by Abcam (1 mentions)
5 protocols using donkey anti goat 594
1
Immunostaining of Cholinergic and Glutamatergic Neurons
2
Immunofluorescence and H&E Analysis of Mouse Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were terminally anesthetized and perfused with PBS followed by 4% PFA. Brains were removed and embedded in paraffin. 5-μm thick coronal sections were deparaffinized and rehydrated in serial ethanol. For immunofluorescence staining, sections were permeabilized with 0.3% TritonX-100 for 15 min, then incubated with blocking solution consisting of 5% donkey serum, followed by incubating with antibodies against GFAP (Abcam), AQP4 (Santa Cruz), Claudin5 (Life Tech) at 4°C overnight. After washing with PBS, slices were incubated with appropriate fluorochrome conjugated secondary antibodies: donkey anti-rabbit 488 (Invitrogen), donkey anti-goat 594 (Invitrogen), donkey anti-rabbit 546 (Invitrogen), respectively, at room temperature for 1 h. Finally, all slices were incubated with fluoro-shield mounting medium with DAPI (Abcam). Images were taken with a fluorescence microscope (model BX-61, Olympus). To get the image with the whole lesion in each slice, we took 10–15 visual fields in a row under a × 40 field of microscope around the lesion site (GFAP and AQP4 loss), then merged these images into a bigger one using the photoshop7.0 software. For H&E Staining, tissue sections were stained with hematoxylin and eosin. Images were taken with a microscope (model BX-61, Olympus). The data were quantified using ImageJ.
3
Characterizing Pluripotency in Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunocytochemistry was used to detect OCT4, SOX2, NANOG, SSEA1, and TRA1 81 in two different passage windows for the three lineages: p16, p15, and p9 for C1, C2, and C3, respectively, and again after p20 (p23, p22, and p22, respectively). The cultured putative piPSCs were fixed in paraformaldehyde for 10 min and washed in PBS. The pluripotency-related markers test was performed as previously described[36 ]. Briefly, the antibodies were used to detect OCT4 (1:100, cat# SC8628, Santa Cruz), SOX2 (1:500, cat# ab97959; Abcam), NANOG (1:100, cat# ab77095, Abcam), SSEA1 (1:50, cat# MAB4301, Millipore) and TRA1 81 (1:50, cat# MAB4381, Millipore), and the respective secondary antibodies were used (donkey anti-goat 594, cat# A11058, donkey anti-rabbit 488, A21206, 1:500, donkey anti-goat 488, cat# A11055, Invitrogen, 1:500 goat anti-mouse 594, cat# A21044, Invitrogen). When necessary, the cells underwent permeabilization and blocking following previously described methods[37 (link)]. At the end of each protocol, the cell nuclei were labelled with Hoechst 33342 (1:1000) and analysed using the EVOS™ photodocumentation system.
4
Immunofluorescent Staining of Mouse Brain Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Coronal sections were air-dried, washed with 1X PBS, blocked with 2% cold water fish skin gelatin (Sigma, Inc., St. Louis, MO, USA) in 0.2% triton-X100 for 1 h, incubated with a primary antibody diluted in blocking solution overnight at room temperature (RT), and then washed with 1X PBS six times 10 min each and incubated with appropriate secondary antibody solution (1:250 dilution) for 1 h at RT. Slides were then washed with 1X PBS and then mounted with media-containing DAPI counterstain (SouthernBiotech, Birmingham, AL, USA). Antibodies used are goat anti-EphA4 at 1:100 dilution (R&D systems, Minneapolis, MN, USA), rabbit anti-TMEM119 at 1:100 dilution (Abcam, Waltham, MA, USA), rabbit anti-CCR2 at 1:100 dilution (Abcam, Waltham, MA, USA), rat anti-CD68 at 1:100 dilution (Thermofisher, USA), rabbit anti-Iba1 at 1:200 dilution (FUJIFILM Wako Chemicals U.S.A. Corporation), donkey anti-goat 594 (Thermofisher, USA), donkey anti-rabbit 647 (Thermofisher, USA), donkey anti-rabbit 555 (Thermofisher, USA), and donkey anti-rat 647 (Thermofisher, USA). Image acquisition was performed using a Zeiss 880 confocal microscope (Carl-Zeiss, Oberkochen, Germany).
5
Immunohistochemical Analysis of Mouse Knee Joints
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sagittal sections from uninjured, 3DPI, and 7DPI knee joints of BL6 mice were used for IHC (n ≥ 3/group). Primary antibodies were incubated overnight at 4 °C in a dark, humid chamber following antigen retrieval. Secondary antibodies were incubated for 2 h at room temperature in a dark, humid chamber at 1:500. Negative control slides were incubated with secondary antibody only. Stained slides were mounted with Prolong Gold with DAPI (Molecular Probes, Eugene, OR, USA). Slides were imaged using a Leica DM5000 microscope (Leica Microsystems, Wetzlar, Germany). ImagePro Plus V7.0 Software, a QIClick CCD camera (QImaging, Surrey, BC, Canada), and ImageJ V1.53 Software were used for imaging and photo editing. Primary antibodies included: CYTL1 (Proteintech, Rosemont, IL, USA; 15856-1-AP (1:75)); MATN3 (R&D, Minneapolis, MN, USA; AF3357 (1:100)); SPP1 (Abcam, Cambridge, UK; ab218237 (1:100)); MMP3 (Abcam, Cambridge, UK; ab52915 (1:100)); CHIL1 (Thermofisher, Waltham, MA, USA; MA5-36122 (1:100)); and INHBA (Thermofisher, Waltham, MA, USA; 10651-1-AP (1:100)). Secondary antibodies included: Chicken anti-rabbit 488 (Thermofisher, Waltham, MA, USA; A21441), Chicken anti-rabbit 594 (Thermofisher, Waltham, MA, USA; 21442), and Donkey anti-goat 594 (Thermofisher, Waltham, MA, USA; A11058).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!