Formaldehyde pbs

Manufactured by Tousimis

Formaldehyde-PBS is a laboratory reagent used for the fixation and preservation of biological samples. It is a solution composed of formaldehyde and phosphate-buffered saline (PBS). Formaldehyde-PBS is commonly used to maintain the structural integrity of cells and tissues during various experimental procedures, such as microscopy, immunohistochemistry, and flow cytometry.

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Lab products found in correlation

Triton x 100, by Bio-Rad (3 mentions) Tween 20, by Bio-Rad (3 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Phosphate buffered saline (pbs), by Corning (3 mentions) In cell analyzer 6000 cell imaging system, by GE Healthcare (2 mentions) Goat serum, by Thermo Fisher Scientific (2 mentions) Phosphate buffered saline (pbs), by Bio-Rad (1 mentions) Clear bottom black 96 well plate, by Corning (1 mentions) Axiovert 200, by Zeiss (1 mentions) Prolong gold reagent, by Thermo Fisher Scientific (1 mentions) Slidebook software, by Olympus (1 mentions) Spinning disc confocal microscope dsu ix81, by Olympus (1 mentions) Zen software, by Zeiss (1 mentions)

Close spelling variants of this product

Formaldehyde

4 protocols using formaldehyde pbs

Imaging Differentiated A152T NPCs

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A152T NPCs were plated and differentiated in 96-well black clear-bottom plates (Corning) POL-coated for 6 weeks. Neurons were fixed with 4% (v/v) formaldehyde-PBS (Tousimis, Rockville, MD) for 20 min, washed in PBS (Corning), incubated in blocking/permeabilization buffer [10 mg/mL BSA (Sigma), 0.05% (v/v) Tween-20 (Biorad), 2% (v/v) goat serum, 0.1% Triton X-100 (Biorad), 92% (v/v) PBS] for 2 hr, followed by overnight incubation with primary antibodies (see antibody section), PBS washed, and then incubated with the corresponding AlexaFluor conjugated secondary antibodies (Life Technologies, Carlsbad, CA). Image acquisition was done with the IN Cell Analyzer 6000 Cell Imaging System (GE Healthcare Life Sciences, Marlborough, MA).

Silva M.C., Ferguson F.M., Cai Q., Donovan K.A., Nandi G., Patnaik D., Zhang T., Huang H.T., Lucente D.E., Dickerson B.C., Mitchison T.J., Fischer E.S., Gray N.S, & Haggarty S.J. (2019). Targeted degradation of aberrant tau in frontotemporal dementia patient-derived neuronal cell models. eLife, 8, e45457.

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Immunostaining of Differentiated Neurons

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NPCs were plated at a starting density of ~ 90,000 cells/cm² in black, clear flat bottom, POL-coated 96-well plates (Corning) in DMEM/F12-B27 media and differentiated for six weeks, followed by compound treatment. Neurons were fixed with 4% (v/v) formaldehyde-PBS (Tousimis) for 30 min, washed in PBS (Corning), incubated in blocking/permeabilization buffer [10 mg/mL BSA (Sigma), 0.05% (v/v) Tween-20 (Bio-Rad), 2% (v/v) goat serum (Life Technologies), 0.1% Triton X-100 (Bio-Rad), in PBS] for 2h, and incubated with primary antibodies overnight (Tau K9JA at 1:1000, MAP2 at 1:1000, Hoechst-33342 at 1:2500). Cells were washed with PBS and incubated with the corresponding AlexaFluor-conjugated secondary antibodies at 1:500 dilution (Life Technologies). Image acquisition was done with a Zeiss AxioVert 200 inverted fluorescence microscope.

Krichevsky A., Nguyen L., Wei Z., Silva M., Barberán-Soler S., Rabinovsky R., Muratore C., Stricker J., Hortman C., Young-Pearse T, & Haggarty S. (2023). Small Molecule Regulators of microRNAs Identified by High-Throughput Screen Coupled with High-Throughput Sequencing. Research Square.

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Immunofluorescence Imaging of Cells

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Cells were seeded in wells of poly-D-lysine coated 96-well glass-bottom
Sensoplate (Greiner Bio-One) and grown to 70% confluency. Cells were fixed for
10 min in 2% formaldehyde/PBS (Tousimis) at 25°C, permeabilized in 0.1%
Triton X-100/PBS and blocked in 4% BSA in PBS. Cells were incubated for 1 h with
the respective primary antibodies followed by incubation (30 min; 25°C)
with species-specific secondary antibody. Cells were mounted on slides using the
Prolong Gold reagent (Life Technologies) containing
4′,6-diamidino-2-phenylindole (DAPI). Confocal microscopy was conducted
using a LSM 710 NLO Zeiss Multiphoton Laser Point scanning confocal microscope
equipped with a multi-photon Mai-Tai laser HB – DeepSee system (690-1024
nm). Images were acquired using ZEN software (Zeiss). Alternatively, a Spinning
Disc confocal microscope DSU-IX81 (Olympus) was used and images were captured
using Slidebook software (Olympus). Images were further processed using ImageJ
(http://www.macbiophotonics.ca) and combined
in Photoshop CC (Adobe System). Signal co-localization was determined using
Coloc2 utility (http://imagej.net/coloc_2).

Chernov A.V., Remacle A.G., Hullugundi S.K., Cieplak P., Angert M., Dolkas J., Shubayev V.I, & Strongin A.Y. (2018). Amino acid sequence conservation of the algesic MBP fragment is required for its interaction with CDK5 and function in pain. The FEBS journal, 285(18), 3485-3502.

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Neuronal Differentiation and Immunostaining

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NPCs were plated at a starting density of 110,000 cells/cm² in black POL-coated 96-well clear-bottom plates (Corning), in DMEM/F12-B27 media and differentiated for six weeks, followed by compound treatment. Neurons were fixed with 4% (v/v) formaldehyde-PBS (Tousimis) for 30 min, washed in PBS (Corning), incubated in blocking/permeabilization buffer [10 mg/mL BSA (Sigma), 0.05% (v/v) Tween-20 (Bio-Rad), 2% (v/v) goat serum (Life Technologies), 0.1% Triton X-100 (Bio-Rad), in PBS] for 2 h, and incubated with primary antibodies overnight (Supplementary Table 2: P-Tau PHF1 1:1000, Tau K9JA 1:1000, MAP2 1:1000, Hoechst 33342 1:2500). Cells were washed with PBS and incubated with the corresponding AlexaFluor-conjugated secondary antibodies at a 1:500 dilution (Life Technologies, Supplementary Table 2,). Image acquisition was done with the InCell Analyzer 6000 Cell Imaging System (GE Healthcare Life Sciences) and micrographs were assembled using Adobe Photoshop 2020 version 21.1.2.

Silva M.C., Nandi G.A., Tentarelli S., Gurrell I.K., Jamier T., Lucente D., Dickerson B.C., Brown D.G., Brandon N.J, & Haggarty S.J. (2020). Prolonged tau clearance and stress vulnerability rescue by pharmacological activation of autophagy in tauopathy neurons. Nature Communications, 11, 3258.

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