Gluteraldehyde

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom

Gluteraldehyde is a chemical compound used as a disinfectant and fixative in various laboratory applications. It is a colorless, oily liquid with a pungent odor. Gluteraldehyde's primary function is to act as a cross-linking agent, effectively preserving and fixing biological specimens for microscopic examination and analysis.

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Lab products found in correlation

Formaldehyde, by Merck Group (3 mentions) Formaldehyde, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) P35g 0 10 c, by MatTek (1 mentions) Sodium deoxycholate, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Dimethylformamide, by Merck Group (1 mentions) Anti actin, by Abcam (1 mentions) Goat anti rabbit igg hrp, by Abcam (1 mentions) Goat anti mouse igg hrp, by Abcam (1 mentions) Anti ku70, by Abcam (1 mentions) β actin, by Abcam (1 mentions) Anti gfp, by Roche (1 mentions) Crystal violet, by Pro-Lab Diagnostics (1 mentions) 5 bromo 4 chloro 3 indolyl β d galactoside x gal, by Promega (1 mentions) Polybrene, by Merck Group (1 mentions) Evo ma10, by Zeiss (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Tcs sp2 system, by Leica (1 mentions)

28 protocols using gluteraldehyde

Evaluating Calcium Mineralization in Saos-2 Cells

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Saos-2 cells were grown and exposed to the Co or Ti nanoparticles changing both the medium and nanoparticles twice a week. After 21 days, the medium was removed from all wells, cells were fixed with 10% (v/v) gluteraldehyde (Sigma Aldrich, UK) in sterile Phosphate Buffer Solution (PBS) (100 µl) for 10 minutes at 37 o C. Then the gluteraldehyde solution was removed and cells were washed three times with PBS.
Calcium was stained with 100 µl of Alizarin red staining (ARS) (Sigma Aldrich, UK) (1 % w/v in distilled water) at 37 o C. After 20 minutes, ARS was removed and the wells washed with Milli-Q water; the staining was dissolved with 100 µl of 10% Acetic acid (Sigma Aldrich, UK). After 30 minutes, 50 µl from each wells were transferred into a 96-well plate and absorbance measurements carried out using a spectrophotometer (ELISA Reader Labtech LT-500MS) at a wavelength of 405nm [24] . All mineralisation tests were performed in triplicates.

Perni S., Yang L., Preedy E.C, & Prokopovich P. (2018). Cobalt and Titanium nanoparticles influence on human osteoblast mitochondrial activity and biophysical properties of their cytoskeleton. Journal of colloid and interface science, 531.

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PALM Imaging of Caenorhabditis elegans

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For PALM, the worms were harvested similarly as in the case of confocal microscopy. They were then fixed in a 4% formaldehyde (Thermo Scientific) or 4% formaldehyde+0.2% gluteraldehyde (Sigma-Aldrich) solution in phosphate-buffered saline (PBS) buffer, the latter resulting in a neglectable fraction of mobile single molecules during PALM imaging63 . The mixture of fixation agents and worms was then incubated on densely Poly-L-lysine (Sigma-Aldrich) coated imaging chambers (P35G-0-10-C; MatTek Corporation) to promote the attachment of worms on the coverslip, probably through the crosslinking of the worm with Poly-L-Lysine. The fixation mixture was removed and the worms were quickly immersed in 50–150 μl CyGel^TM (BioCompare) to immobilise them, match the refractive index and prevent the samples from drying out. The outer ring of the imaging dish was filled with approximately 1 ml 1-2% agarose as to further prevent drying of the sample by saturating the vapour pressure inside the chamber.

Vangindertael J., Beets I., Rocha S., Dedecker P., Schoofs L., Vanhoorelbeeke K., Hofkens J, & Mizuno H. (2015). Super-resolution mapping of glutamate receptors in C. elegans by confocal correlated PALM. Scientific Reports, 5, 13532.

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Corneal β-Galactosidase Visualization

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Patterns of β-galactosidase activity were observed in whole corneas of XLacZ^Tg/− hemizygous female animals after staining with 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-Gal) substrate. Eyeballs were enucleated from 15 to 20 week old mice and rinsed briefly in PBS. Eyes were placed in fix solution for 1 h at 4°C (2% formaldehyde (Sigma), 0.2% gluteraldehyde (Sigma), 2 mM MgCl₂, 5 mM EGTA in 0.1 M phosphate buffer pH 7.4) and then washed three times for 20 min with in excess of detergent wash (0.01% sodium deoxycholate (Sigma, Life Sciences), 0.02% Nonidet P40 (NP40 substitute, BioChemika, Fluka 74 385), 0.05% BSA (Sigma) in 0.1 M phosphate buffer). Eyes were then placed in X-Gal staining solution (25 ml detergent wash, 0.085% NaCl, 5 mM K₃Fe(CN)₆, 5 mM K₄Fe(CN)₆, 0.1% X-Gal dissolved in 0.5 ml dimethyl formamide (Sigma)) overnight at 37°C. After X-Gal staining, eyes were fixed in 4% paraformaldehyde for 1 h and washed with PBS before taking them up through an ethanol series (1 × 50%, 2 × 70% for 20 min each) and stored at 4°C. Corneas were dissected from the eyes as previously described, and images were obtained under a stereomicroscope with the cornea facing uppermost.

Findlay A.S., Panzica D.A., Walczysko P., Holt A.B., Henderson D.J., West J.D., Rajnicek A.M, & Collinson J.M. (2016). The core planar cell polarity gene, Vangl2, directs adult corneal epithelial cell alignment and migration. Royal Society Open Science, 3(10), 160658.

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GFP-nCLU and IPO13 Protein Interaction in Oxidative Stress

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HeLa cells co-expressing GFP or GFP-nCLU proteins with DsRed2-IPO13 were treated with or without 125 μM H₂O₂ (Sigma, St. Louis, MO, USA) for 1 h prior to cell lysis. HeLa cells expressing GFP or GFP-IPO13 protein treated with or without 125 μM H₂O₂ (Sigma, St. Louis, MO, USA) for 1 h underwent protein cross-link using Gluteraldehyde (Sigma, St. Louis, MO, USA) as previously described [46 (link)], after which they were lysed immediately. In both cases, immunocomplexes were precipitated and eluted using GFP-Trap resin (ChromoTek, Proteintech, Rosemont, IL, USA) according to the manufacturer’s instructions. Input and immunoprecipitation fractions were analysed by Western analysis using anti-IPO13 (Cat no. 11696-2-AP, Protein Tech, Rosemont, IL, USA), anti-KU70 (Cat no. ab3114, Abcam, Cambridge, UK), anti-GFP (Cat no. 11814460001, Roche, Basel, Switzerland), anti-Actin (Cat no. ab3280 Abcam, Cambridge, UK), and β-actin (CST, Cat no. 3700) primary antibodies and the secondary antibodies goat anti-mouse IgG HRP (AP308P) and goat anti-rabbit IgG HRP (AP307P).

Gajewska K.A., Jans D.A, & Wagstaff K.M. (2023). The Nuclear Transporter Importin 13 Can Regulate Stress-Induced Cell Death through the Clusterin/KU70 Axis. Cells, 12(2), 279.

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Cell Migration Assay Protocol

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Cells were seeded at a density of 5 × 10⁴ cells in the upper compartment of migration inserts (Becton Dickinson, Franklin Lakes, NJ, USA). The lower chamber was filled with media containing fibroblast-conditioned media. Cells were incubated for 48 (MDA-MB-231) or 72 h (BT20 and MDA-MB-453). Following incubation, non-migrated cells were removed using a PBS-soaked cotton swab. In all, 1% gluteraldehyde (Sigma, St Louis, MO, USA) was used to fix the migrated cells, followed by staining with 0.1% crystal violet (Pro-Lab Diagnostics, Chesire, UK). Migrated cells were counted using a light microscope at a magnification of × 20. Five fields were counted in each well and averaged. The percentage of migrated cells was calculated compared with scrambled or vehicle control cells. Results are shown as mean of independent experiments carried out in triplicate±s.e.m.

Mullooly M., McGowan P.M., Kennedy S.A., Madden S.F., Crown J., O' Donovan N, & Duffy M.J. (2015). ADAM10: a new player in breast cancer progression?. British Journal of Cancer, 113(6), 945-951.

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Titration of Murine Leukemia Virus

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NIH3T3 cells (500,000) were seeded in 5 cm² plates in 1 ml DMEM, for 24 hr and then infected with 10⁻³–10⁻⁷ dilutions of MulV in 1 ml DMEM, with 4 μg/ml Polybrene (sigma Aldrich) and no fetal calf serum (FCS). Two hours postinfection, 10% FCS and DMEM were added to achieve a total volume of 5 ml, and cells were incubated for a further 48 hr at 37°C in 5% CO₂. After 48 hr, NIH3T3 cells were fixed using 0.2% gluteraldehyde (Sigma Aldrich), 2% formaldehyde (Sigma Aldrich), 2 mM MgCl₂ in 1× PBS, for 10 min at room temperature and stained with 0.1% 5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐galactoside (X‐Gal, Promega). The number of blue cells was counted in five 1 mm² randomly selected squares and the titer of the virus was calculated based on the plate area and the dilution of the virus.

Matza‐Porges S, & Nathan D. (2017). A biosafety level 2 virology lab for biotechnology undergraduates. Biochemistry and Molecular Biology Education, 45(6), 537-543.

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Silanization and Gluteraldehyde Fixation of Glass Coverslips

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Glass coverslips were coated with a drop of 3-aminopropyltrimethoxysilane spread evenly on the surface. Each coverslip was then washed with deionized water, placed into foil packets, and autoclaved. Once autoclaved, the coverslips were transferred to a sterile petri dish containing 0.5% gluteraldehyde (70% stock solution, Sigma) in PBS and incubated for 30 minutes at room temperature. This was followed by 5, 10 minute washes in autoclaved deionized water. After the last wash, the coverslips were laid out onto a piece of autoclaved foil in a biosafety cabinet to dry.

Davis J.T, & Foster W.J. (2018). Substrate Stiffness Influences the Time Dependence of CTGF Protein Expression in Müller Cells. International physiology journal, 1(1), 1.

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Scanning Electron Microscopy of Promastigotes

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Control and treated promastigotes (2 × 10⁶ cells) were fixed with 2.5% gluteraldehyde (Sigma Aldrich), dehydrated in ethanol, critical point-dried in CO₂, mounted on stubs, sputtered with a thin gold layer5 , 6 and observed under a scanning electron microscope (Model: ZEISS EVO-MA 10).

Mukherjee D., Singh C.B., Dey S., Mandal S., Ghosh J., Mallick S., Hussain A., Swapana N., Ross S.A, & Pal C. (2015). Induction of apoptosis by zerumbone isolated from Zingiber zerumbet (L.) Smith in protozoan parasite Leishmania donovani due to oxidative stress. The Brazilian Journal of Infectious Diseases, 20(1), 48-55.

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Virus Quantification by Flow Cytometry

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Duplicate aliquots (1.8 ml) of each sample were dispensed into 2 ml-cryotubes containing gluteraldehyde (0.5% final concentration, electron microscopy grade, Sigma-Aldrich). Samples were fixed at 4^∘C for 30 min then immediately frozen at -80^∘C, before being analyzed using FC within 4–6 weeks (see below). Samples were shipped frozen on dry-ice back to Oregon State University (OSU). Due to logistic constraints, the samples could not be flash-frozen in liquid nitrogen before being stored at -80^∘C. This is known to account for some virus losses, however they are reported to be minimal (<10%; Brussaard, 2004a (link)). We therefore expect that virus loss in our samples was minimal, and that our data represent conservative estimates of VA. FC analysis of viruses, heterotrophic bacteria and phytoplankton were performed on a Becton Dickinson (BD) FACSCalibur flow cytometer (15 mW argon laser exciting at 488 nm, BD, San Jose, CA, USA), as described below.

Payet J.P., McMinds R., Burkepile D.E, & Vega Thurber R.L. (2014). Unprecedented evidence for high viral abundance and lytic activity in coral reef waters of the South Pacific Ocean. Frontiers in Microbiology, 5, 493.

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Invadopodia Formation and Degradation Assay

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Invadopodia formation assays were performed as previously described [53 (link)]. Briefly, coverslips were coated with 50 µg/ml poly-l-lysine (Sigma), followed by 0.5% gluteraldehyde (sigma) and inverted on an 80 ul drop of unlabeled, Alexa488-labeled,or Alexa594-labled, incubated with 5 mg/ml NaBorohydride (Sigma), and washed extensively with PBS. Cells were serum starved for 6–12 h, lifted and plated on coverslips with 50 um GM6001 for 3 h (T0), followed by extensive washes with PBS, incubated for 1 (T1) or 2 h (T2), as indicated, fixed, permeabilized and stained with indicated antibodies and phalloidin. GABA (1 mM), EGF (100 ng/ml) or Erlotinib (20 µM) was added, as indicated, during cell plating for 4-hrs. For analysis of invadopodia degradation and actin core size samples were imaged using a 63 × (NA 1.4) lens on a Leica DM-IRE2 upright microscope with a Leica TCS SP2 system (Leica, Heidelberg, Germany) and areas of degradation per cell or actin cores were analyzed using ImageJ software Analyze Particle program. For Western blot analysis, cells were plated on unlabeled gelatin and treated as above followed by lysis at T0, T1, and T2.

Williams K.C., Cepeda M.A., Javed S., Searle K., Parkins K.M., Makela A.V., Hamilton A.M., Soukhtehzari S., Kim Y., Tuck A.B., Ronald J.A., Foster P.J., Chambers A.F, & Leong H.S. (2019). Invadopodia are chemosensing protrusions that guide cancer cell extravasation to promote brain tropism in metastasis. Oncogene, 38(19), 3598-3615.

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