Lung and FALC were prepared by fixation in 4% paraformaldehyde at 4 °C for 1 h, incubating in 20% sucrose overnight for cryoprotection, and embedding in 15% sucrose/7.5% gelatin diluted in PBS. Overall, 12 μm frozen sections were cut using a Leica CM 3050 S cryostat. Sections were pre-blocked/permeabilized in PBS containing 3% BSA and 0.05% Triton X-100 and stained with primary antibodies anti-mCherry (M11217, Invitrogen), anti-Relmα (500-P214, PeproTech), or anti-actin α-smooth muscle FITC conjugated (1A4, Sigma), followed by secondary antibodies anti-rat IgG Alexa Fluor 568 (A11077, Life Technologies) and anti-rabbit IgG Alexa Fluor 488 (A21206, Life Technologies) for 1 h at the room temperature, or directly conjugated CD3 (clone 145-2C11), B220 (clone RA3-6B2), and CD169 (clone 3D6.112). Image acquisition was performed using either a Zeiss LSM 780 confocal microscope and ZEN 2010 acquisition software or a Nikon HCA fluorescence microscope and NIS-Elements 4.30.01 acquisition software for live cell imaging. Adobe Photoshop 11.0.2 was used to adjust brightness, contrast and colour balance (changes were applied to all images in equal measure).
Anti relmα 500 p214
Manufactured by Thermo Fisher Scientific
Anti-Relmα (500-P214) is a laboratory reagent produced by Thermo Fisher Scientific. It functions as an antibody that specifically recognizes and binds to the Relmα protein. This reagent is intended for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti rat igg alexa fluor 568, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Cm3050s cryostat, by Leica (2 mentions)
Lsm 780 confocal microscope, by Nikon (1 mentions)
Anti mcherry, by Thermo Fisher Scientific (1 mentions)
M11217, by Thermo Fisher Scientific (1 mentions)
Lsm 780 confocal microscope, by ZenBio (1 mentions)
2 protocols using anti relmα 500 p214
1
Cryoprotection and Immunofluorescence Imaging
2
Fixation, Cryosectioning, and Immunostaining
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Lung and FALC were prepared by fixation in 4% paraformaldehyde at 4 °C for 1 h, incubating in 20% sucrose overnight for cryoprotection, and embedding in 15% sucrose/7.5% gelatin diluted in PBS. Overall, 12 μm frozen sections were cut using a Leica CM 3050 S cryostat. Sections were pre-blocked/permeabilized in PBS containing 3% BSA and 0.05% Triton X-100 and stained with primary antibodies anti-mCherry (M11217, Invitrogen), anti-Relmα (500-P214, PeproTech), or anti-actin α-smooth muscle FITC conjugated (1A4, Sigma), followed by secondary antibodies anti-rat IgG Alexa Fluor 568 (A11077, Life Technologies) and anti-rabbit IgG Alexa Fluor 488 (A21206, Life Technologies) for 1 h at the room temperature, or directly conjugated CD3 (clone 145-2C11), B220 (clone RA3-6B2), and CD169 (clone 3D6.112). Image acquisition was performed using either a Zeiss LSM 780 confocal microscope and ZEN 2010 acquisition software or a Nikon HCA fluorescence microscope and NIS-Elements 4.30.01 acquisition software for live cell imaging. Adobe Photoshop 11.0.2 was used to adjust brightness, contrast and colour balance (changes were applied to all images in equal measure).
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