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permeabilization buffer a  (Applichem inc)
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Applichem inc permeabilization buffer a
Alkyne lipid imaging is compatible with immunofluorescence and commonly used cellular stains. MEF cells were treated for 24 h with 1 µM alkyne-oleate and fixed for 10 min. A: Prior treatment with saponin for membrane <t>permeabilization</t> did not diminish or change subsequent click-labeling of cellular structures. Cells were treated for 2 h with 0%, 0.1%, or 0.01% saponin before subjecting them to the click reaction using 2 mM CuTFB and 10 µM ASBDP. B: Immunofluorescence labeling is preserved during the CuAAC click procedure. Cells were first subjected to immunofluorescence staining for commonly used organelle marker proteins of three different cellular compartments: endoplasmic reticulum (PDI), mitochondria (Tom20), and lysosomes (LAMP1). Subsequently, cells were subjected to the click reaction using 2 mM CuTFB and 10 µM ASBDP. C: Click labeling of alkyne lipids by CuAAC (using 2 mM CuTFB and 10 µM ASBDP) is compatible with commonly used fluorescence stains. Cells were stained in situ using mitotracker CMXRos (50 nM, 15 min) before fixation and subsequent click labeling or stained with LD540 or DAPI after the click labeling. All images were acquired using the Orca-Flash4.0 camera and are shown at the optimal display level each. Scale bars, 10 μm.
Permeabilization Buffer A, supplied by Applichem inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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permeabilization buffer a - by Bioz Stars, 2026-10
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Alkyne lipid imaging is compatible with immunofluorescence and commonly used cellular stains. MEF cells were treated for 24 h with 1 µM alkyne-oleate and fixed for 10 min. A: Prior treatment with saponin for membrane permeabilization did not diminish or change subsequent click-labeling of cellular structures. Cells were treated for 2 h with 0%, 0.1%, or 0.01% saponin before subjecting them to the click reaction using 2 mM CuTFB and 10 µM ASBDP. B: Immunofluorescence labeling is preserved during the CuAAC click procedure. Cells were first subjected to immunofluorescence staining for commonly used organelle marker proteins of three different cellular compartments: endoplasmic reticulum (PDI), mitochondria (Tom20), and lysosomes (LAMP1). Subsequently, cells were subjected to the click reaction using 2 mM CuTFB and 10 µM ASBDP. C: Click labeling of alkyne lipids by CuAAC (using 2 mM CuTFB and 10 µM ASBDP) is compatible with commonly used fluorescence stains. Cells were stained in situ using mitotracker CMXRos (50 nM, 15 min) before fixation and subsequent click labeling or stained with LD540 or DAPI after the click labeling. All images were acquired using the Orca-Flash4.0 camera and are shown at the optimal display level each. Scale bars, 10 μm.

Journal: Journal of Lipid Research

Article Title: A highly sensitive protocol for microscopy of alkyne lipids and fluorescently tagged or immunostained proteins [S]

doi: 10.1194/jlr.D070565

Figure Lengend Snippet: Alkyne lipid imaging is compatible with immunofluorescence and commonly used cellular stains. MEF cells were treated for 24 h with 1 µM alkyne-oleate and fixed for 10 min. A: Prior treatment with saponin for membrane permeabilization did not diminish or change subsequent click-labeling of cellular structures. Cells were treated for 2 h with 0%, 0.1%, or 0.01% saponin before subjecting them to the click reaction using 2 mM CuTFB and 10 µM ASBDP. B: Immunofluorescence labeling is preserved during the CuAAC click procedure. Cells were first subjected to immunofluorescence staining for commonly used organelle marker proteins of three different cellular compartments: endoplasmic reticulum (PDI), mitochondria (Tom20), and lysosomes (LAMP1). Subsequently, cells were subjected to the click reaction using 2 mM CuTFB and 10 µM ASBDP. C: Click labeling of alkyne lipids by CuAAC (using 2 mM CuTFB and 10 µM ASBDP) is compatible with commonly used fluorescence stains. Cells were stained in situ using mitotracker CMXRos (50 nM, 15 min) before fixation and subsequent click labeling or stained with LD540 or DAPI after the click labeling. All images were acquired using the Orca-Flash4.0 camera and are shown at the optimal display level each. Scale bars, 10 μm.

Article Snippet: In detail, cells were permeabilized for 15 min in permeabilization buffer A [PBS/1% cold fish gelantine/0.01% saponin (Applichem #A2542)] for permeabilization of plasma membrane only or permeabilization buffer B (PBS/1% cold fish gelantine/0.1% saponin) for permeabilization of all cellular membranes.

Techniques: Imaging, Immunofluorescence, Membrane, Labeling, Staining, Marker, Fluorescence, In Situ