Canine platelets were incubated on an ITS surface for 30 min and then fixed with 4% paraformaldehyde plus 0.5% Triton X in PBS for 30 min. After washing with PBS three times, the platelets were immersed in 2% BSA (bovine serum albumin) for 1 hour at room temperature. 2.5 μg/ml vinculin antibody (FAK100, Millipore, mouse anti-Vinculin) was added to platelet surface and incubated for 1 hour. After washing, 2.5 μg/ml secondary antibody (AP192SA6, Millipore) was added to the platelet surface and incubated for 1 hour at room temperature. After washing, the ITS force map and vinculin were co-imaged with the fluorescence microscope.
Ap192sa6
Manufactured by Merck Group
Sourced in United States
The AP192SA6 is a laboratory equipment product. It is a device used for scientific analysis and research purposes. The core function of this product is to perform specific tasks within a laboratory setting. No further details about the intended use or application of this product can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
5 protocols using ap192sa6
1
Visualizing Platelet Vinculin Dynamics
2
Visualizing Membrane-bound DNase X in MDA-MB-231 Cells
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MDA‐MB‐231 cells were plated on an SNS‐coated glass surface and incubated at 37 °C for 2 h. Next, the cells were fixed with 4% paraformaldehyde for 20 min and blocked with 3% BSA for 1 h, followed by the staining of primary antibody anti‐DNase X (H00001774‐M02, Abnova, Taiwan) with 1:100 dilution. The cells were then treated with secondary antibody (AP192SA6, Millipore, MA, USA) with 1:200 dilution for 1 h, Immunostained cell samples were observed under Nikon Ti‐E microscope, with Cy3 channel for SNS signal and GFP channel for DNase X staining. During the immunostaining process, the cell membrane was not permeablized to avoid staining cytosolic DNase X that may cloud the imaging of membrane‐bound DNase X.
3
Visualizing Membrane-bound DNase X in MDA-MB-231 Cells
4
Visualization of Inhibitory Neurons in Mouse V1
5
Visualization of Inhibitory Neurons in Mouse V1
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Two weeks after the rabies virus injection, mice were anaesthetised with isoflurane, injected with sodium pentobarbital (0.01 mL/g) and perfused trans-cardially with ice-cold phosphate buffered saline (PBS), followed by 4% ice-cold PFA in PBS. The brain was extracted, postfixed for 24h in 4% PFA PBS at 4°C, and finally transferred to a 30% sucrose PBS at 4°C. 50 μm thick coronal sections were serially cut on a vibratome to span the whole V1, washed in PBS, blocked with a 0.3% triton-X100, 1% BSA solution in PBS and processed with the following antibodies: mouse anti-GAD67 primary antibody (1:1500, Millipore MAB5406), donkey anti-mouse IgG Alexa 647 conjugate secondary antibody (1:800, Merk Millipore AP192SA6). Sections were mounted on SuperFrost slides (Molecular Probes), air dried, and cover-slipped with Fluoromount (Sigma-Aldrich). Z-stacks tile scans of each slide were imaged with a Nikon Eclipse Ti2-E inverted epifluorescence microscope and manually aligned to the corresponding sections from in vivo z-stacks. Images were high-pass filtered for display (Extended Data Figure 5 ).
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