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Digitonin

Digitonin is a naturally occurring saponin compound derived from the plant Digitalis purpurea.
It is commonly used as a detergent and permeabilizing agent in biochemical and cell biology research, particularly for the study of membrane proteins and organellar structures.
Digitonin interacts with cholesterol in cell membranes, causing them to become more permeable and allowing for the extraction or fractionation of cellular components.
It is a valuable tool for investigating subcellular organization, protein-protein interactions, and the function of membrane-bound proteins.
Researchers should carefully optimize Digitonin concentrations and incubation times to preserve cellular integrity and target specificity when using this powerful reagent.

Most cited protocols related to «Digitonin»

Nuclei Isolation for ATAC-Seq Analysis

Cited 196 times

See Supplementary
Protocol 2 for a detailed protocol. This protocol is highly similar
to the INTACT method^{19 (link)} and
either protocol can be used for the isolation of nuclei with equivalent results.
All of the steps were carried out at 4 °C. A frozen tissue fragment ~20
mg was placed into a pre-chilled 2-ml Dounce homogenizer containing 2 ml of cold
1× homogenization buffer (320 mM sucrose, 0.1 mM EDTA, 0.1%
NP40, 5 mM CaCl₂, 3 mM Mg(Ac)₂, 10 mM Tris pH 7.8,
1× protease inhibitors (Roche, cOmplete), and 167 μM
β-mercaptoethanol, in water). Tissue was homogenized with approximately
ten strokes with the loose ‘A’ pestle, followed by 20 strokes
with the tight ‘B’ pestle. Connective tissue and residual debris
were precleared by filtration through an 80-μm nylon mesh filter
followed by centrifugation for 1 min at 100 r.c.f. While avoiding the pelleted
debris, 400 μl was transferred to a pre-chilled 2-ml round bottom
Lo-Bind Eppendorf tube. An equal volume (400 μl) of a 50%
iodixanol solution (50% iodixanol in 1× homogenization buffer)
was added and mixed by pipetting to make a final concentration of 25%
iodixanol. 600 μl of a 29% iodixanol solution (29%
iodixanol in 1× homogenization buffer containing 480 mM sucrose) was
layered underneath the 25% iodixanol mixture. A clearly defined
interface should be visible. In a similar fashion, 600 μl of a
35% iodixanol solution (35% iodixanol in 1×
homogenization containing 480 mM sucrose) was layered underneath the 29%
iodixanol solution. Again, a clearly defined interface should be visible between
all three layers. In a swinging-bucket centrifuge, nuclei were centrifuged for
20 min at 3,000 r.c.f. After centrifugation, the nuclei were present at the
interface of the 29% and 35% iodixanol solutions. This band with
the nuclei was collected in a 300 μl volume and transferred to a
pre-chilled tube. Nuclei were counted after addition of trypan blue, which
stains all nuclei due to membrane permeabilization from freezing. 50,000 counted
nuclei were then transferred to a tube containing 1 ml of ATAC-seq RSB with
0.1% Tween-20. Nuclei were pelleted by centrifugation at 500 r.c.f. for
10 min in a pre-chilled (4 °C) fixed-angle centrifuge. Supernatant was
removed using the two pipetting steps described above. Because the nuclei were
already permeabilized, no lysis step was performed, and the transposition mix
(25 μl 2× TD buffer, 2.5 μl transposase (100 nM final),
16.5 μl PBS, 0.5 μl 1% digitonin, 0.5 μl
10% Tween-20, 5 μl water) was added directly to the nuclear
pellet and mixed by pipetting up and down six times. Transposition reactions
were incubated at 37 °C for 30 min in a thermomixer with shaking at
1,000 r.p.m. Reactions were cleaned up with Zymo DNA Clean and Concentrator 5
columns. The remainder of the ATAC-seq library preparation was performed as
described previously¹⁸.

Corces M.R., Trevino A.E., Hamilton E.G., Greenside P.G., Sinnott-Armstrong N.A., Vesuna S., Satpathy A.T., Rubin A.J., Montine K.S., Wu B., Kathiria A., Cho S.W., Mumbach M.R., Carter A.C., Kasowski M., Orloff L.A., Risca V.I., Kundaje A., Khavari P.A., Montine T.J., Greenleaf W.J, & Chang H.Y. (2017). An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues. Nature methods, 14(10), 959-962.

Publication 2017

2-Mercaptoethanol ATAC-Seq Buffers Cell Nucleus Centrifugation Cerebrovascular Accident Connective Tissue Digitonin DNA Library Edetic Acid Filtration iodixanol isolation Nylons Protease Inhibitors Sucrose Tissue, Membrane Tissues Transposase Tromethamine Trypan Blue Tween 20

ATAC-seq Cell Preparation Protocol

Cited 151 times

GM12878 cells were counted five times using a manual hemocytometer. The
mean cell count was used to resuspend the cells to a concentration of 500 cells
per 100 μl by the addition of PBS. From this diluted cell mixture, 100
μl (500 cells) were deposited into a 0.5-ml DNA LoBind tube (Eppendorf
#022431005) containing 400 μl of cold ATAC-seq RSB. This was
done to simulate a work-flow involving FACS sorting. These tubes were
centrifuged at 500 r.c.f. for 10 min in a pre-chilled (4 °C) fixed-angle
centrifuge with 0.6-ml tube adapters. All of the supernatant was removed using
the two pipetting steps described above, first by removing 400 μl with a
P1000 pipette tip followed by removal of the remaining volume with a P200
pipette tip. We note that a gradual but constant removal of supernatant is
crucial and that the final supernatant removal step should be completed in a
single motion to avoid disrupting the cell pellet. After supernatant removal,
lysis and transposition were performed simultaneously to avoid cell loss, and
the total reaction volume was reduced for the same reason. As such, 10
μl of transposition mix (3.3 μl PBS, 1.15 μl water, 5
μl 2× TD Buffer, 0.25 μl 1:10 diluted Tn5
enzyme^{26 (link)}, 0.1
μl 1% digitonin, 0.1 μl 10% Tween-20, and 0.1
μl 10% NP40) was added directly to the invisible cell pellet,
and the pellet was resuspended by pipetting up and down six times. The
transposition reaction was incubated at 37 °C for 30 min in a
thermomixer with shaking at 1,000 r.p.m. Note that Tn5 should be diluted in
1× TD Buffer (for example, 5 μl 2× TD Buffer, 4
μl of water, 1 μl Tn5).

Publication 2017

ATAC-Seq Buffers Cells Cold Temperature Digitonin F 500 Tween 20

Fast-ATAC Sequencing of Sorted Cells

Cited 82 times

This protocol has been optimized for blood cells. We note that digitonin is a gentle detergent and this protocol may not be ideal for cell lines and other cell types that are more resistant to lysis. 5,000 sorted cells in FACS Buffer were pelleted by centrifugation at 500 RCF for 5 minutes at 4C in a pre-cooled fixed-angle centrifuge. All supernatant was removed using two pipetting steps being careful to not disturb the not visible cell pellet. 50 ul transposase mixture (25 ul of 2x TD buffer, 2.5 ul of TDE1, 0.5 ul of 1% digitonin, 22 ul of nuclease-free water) (Cat# FC-121-1030, Illumina; Cat# G9441, Promega) was added to the cells and the pellet was disrupted by pipetting. Transposition reactions were incubated at 37°C for 30 minutes in an Eppendorf ThermoMixer with agitation at 300 RPM. Transposed DNA was purified using a QIAgen MinElute Reaction Cleanup kit (Cat# 28204) and purified DNA was eluted in 10 ul elution buffer (10 mM Tris-HCl, pH 8). Transposed fragments were amplified and purified as described previously^{48 (link)} with modified primers^{23 (link)}. Libraries were quantified using qPCR prior to sequencing. All Fast-ATAC libraries were sequenced using paired-end, dual-index sequencing on a NextSeq with 76×8×8×76 cycle reads.

Corces M.R., Buenrostro J.D., Wu B., Greenside P.G., Chan S.M., Koenig J.L., Snyder M.P., Pritchard J.K., Kundaje A., Greenleaf W.J., Majeti R, & Chang H.Y. (2016). Lineage-specific and single cell chromatin accessibility charts human hematopoiesis and leukemia evolution. Nature genetics, 48(10), 1193-1203.

Publication 2016

Blood Cells Buffers Cell Lines Cells Centrifugation Detergents Digitonin Promega Transposase Tromethamine XCL1 protein, human

ATAC-Seq Library Preparation for Primary Human T Cells

Cited 51 times

See Protocol Exchange¹⁷ or Supplementary Protocols 1 and 2 for
a detailed protocol. Cells grown in tissue culture were pretreated with 200 U/ml
DNase (Worthington) for 30 min at 37 °C to remove free-floating DNA and
to digest DNA from dead cells. This medium was then washed out, and the cells
were resuspended in cold PBS. For primary human T cells, cells were sorted using
a Becton Dickinson FACS Aria II instrument based on the expression of CD45, CD3,
and CD4, as described previously^{11 (link)}. After the cells were counted, 50,000 cells were
resuspended in 1 ml of cold ATAC-seq resuspension buffer (RSB; 10 mM Tris-HCl pH
7.4, 10 mM NaCl, and 3 mM MgCl₂ in water). Cells were centrifuged at
500 r.c.f. for 5 min in a pre-chilled (4 °C) fixed-angle centrifuge.
After centrifugation, 900 μl of supernatant was aspirated, which left
100 μl of supernatant. This remaining 100 μl of supernatant was
carefully aspirated by pipetting with a P200 pipette tip to avoid the cell
pellet. Cell pellets were then resuspended in 50 μl of ATAC-seq RSB
containing 0.1% NP40, 0.1% Tween-20, and 0.01% digitonin
by pipetting up and down three times. This cell lysis reaction was incubated on
ice for 3 min. After lysis, 1 ml of ATAC-seq RSB containing 0.1%
Tween-20 (without NP40 or digitonin) was added, and the tubes were inverted to
mix. Nuclei were then centrifuged for 10 min at 500 r.c.f. in a pre-chilled (4
°C) fixed-angle centrifuge. Supernatant was removed with two pipetting
steps, as described before, and nuclei were resuspended in 50 μl of
transposition mix (25 μl 2× TD buffer (recipe in Supplementary Protocol 1) 2.5
μl transposase^{26 (link)} (100
nM final), 16.5 μl PBS, 0.5 μl 1% digitonin, 0.5
μl 10% Tween-20, and 5 μl water) by pipetting up and
down six times. Transposition reactions were incubated at 37 °C for 30
min in a thermomixer with shaking at 1,000 r.p.m. Reactions were cleaned up with
Zymo DNA Clean and Concentrator 5 columns. The remainder of the ATAC-seq library
preparation was performed as described previously¹⁸. All libraries were amplified with a
target concentration of 20 μl at 4 nM, which is equivalent to 80
femtomoles of product. Minor protocol modifications were used for Omni-ATAC in
frozen tissues and in limiting cell numbers. These modifications are outlined in
the corresponding Online Methods sections.

Publication 2017

ATAC-Seq Buffers Cell Nucleus Cells Centrifugation Cold Temperature Digitonin Homo sapiens Magnesium Chloride NRG1 protein, human Pellets, Drug Sodium Chloride T-Lymphocyte Tissues Tromethamine Tween 20 XCL1 protein, human

CUT&RUN Profiling of Transcription Factors and Histone Modifications

Cited 41 times

CUT&RUN was performed as previously described [10 (link)]. Briefly, cells were washed with Wash Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM spermidine and one Roche Complete protein inhibitor tablet per 50 mL), bound to Concanavalin A-coated magnetic beads and incubated with primary antibody diluted in wash buffer containing 0.05% digitonin (Dig Wash) overnight at 4 °C. Cells were then washed and incubated with protein A-MNase (pA-MN) for 1 h at 4 °C. Slurry was washed again and placed on an ice-cold block and incubated with Dig Wash containing 2 mM CaCl₂ to activate pA-MN digestion. After digestion for 30 min, one volume of 2× stop buffer (340 mM NaCl, 20 mM EDTA, 4 mM EGTA, 0.05% Digitonin, 0.05 mg/mL glycogen, 5 µg/mL RNase A, 2 pg/mL heterologous spike-in DNA) was added to stop the reaction, and fragments were released by 30-min incubation at 37 °C. Samples were centrifuged 5 min at 16,000×g, and supernatant was recovered and DNA extracted via phenol–chloroform extraction and ethanol precipitation. Resulting DNA was used as input for library preparation as previously described [10 (link)]. Antibodies used for CUT&RUN in this study were as follows: rabbit anti-Sox2 (Abcam ab92494); rabbit anti-FoxA2 (Millipore 07-633); Guinea-Pig anti-rabbit IgG (antibodies online ABIN101961); rabbit anti-H3K4me2 (Millipore 07-030); rabbit anti-H3K4me3 (Active Motif 39159); rabbit anti-H3K27me3 (Cell Signaling Technologies CST9733); and rabbit anti-CTCF (Millipore 07-729).

Meers M.P., Tenenbaum D, & Henikoff S. (2019). Peak calling by Sparse Enrichment Analysis for CUT&RUN chromatin profiling. Epigenetics & Chromatin, 12, 42.

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Publication 2019

anti-IgG Antibodies Buffers Cavia porcellus Cells Chloroform Cold Temperature Concanavalin A CTGF protein, human Digestion Digitonin DNA Library Edetic Acid Egtazic Acid Ethanol Glycogen HEPES histone H3 trimethyl Lys4 Immunoglobulins Phenol Protein Digestion Proteins Rabbits Ribonucleases Sodium Chloride SOX2 protein, human Spermidine Staphylococcal Protein A Tablet

Most recents protocols related to «Digitonin»

Intracellular Cholesterol and Mitochondrial GSH Analysis

Cholesterol levels were determined fluorometrically using the Amplex Red Cholesterol Assay kit (Thermo Fisher Sci.; A12216). Samples (0.5 × 10⁵ cells) were extracted with chloroform:isopropanol:IGEPAL CA-630 (7:11:0.1) and centrifuged at 13,000 g for 10 min to remove insoluble material. The organic phase was dried under vacuum, dissolved in 1 × cholesterol reaction buffer, and analyzed following the guidelines provided by the supplier. To evaluate the intracellular distribution of the cholesterol load, cells were fixed in 4% paraformaldehyde (PFA) for 20 min at room temperature, and after washing three times, were incubated with the naturally fluorescent polyene antibiotic filipin III (0.25 mg/ml; Sigma-Aldrich, F4767) for 30 min. Images were acquired with a Zeiss Axiophot fluorescence microscope using a 40 × /1.3 N.A. objective. The ImageJ software [34 (link)] was used to calculate the corrected total cell fluorescence (CTCF) by applying the following formula: CTCF = Integrated Density − (Area of selected cell × Mean fluorescence of background readings). Mitochondria from SH-SY5Y cells were isolated by digitonin fractionation as described previously [35 ] and the GSH content was analyzed using the Glutathione Assay Kit (Sigma-Aldrich, CS0260-1 KT) following the manufacturer's instructions.

de Dios C., Abadin X., Roca-Agujetas V., Jimenez-Martinez M., Morales A., Trullas R., Mari M, & Colell A. (2023). Inflammasome activation under high cholesterol load triggers a protective microglial phenotype while promoting neuronal pyroptosis. Translational Neurodegeneration, 12, 10.

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Publication 2023

Antibiotics Biological Assay Buffers Cells Chloroform Cholesterol CTCF protein, human Digitonin Filipin Fluorescence Fractionation, Chemical Glutathione Igepal CA-630 Isopropyl Alcohol Microscopy, Fluorescence Mitochondria paraform Polyenes Protoplasm Vacuum

Intracellular Cholesterol and Mitochondrial GSH Analysis

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Publication 2023

Proteinase K Sensitivity of Yeast Mitochondria

Proteinase K protection assays were performed as previously described (70 (link), 71 (link)). Forty micrograms yeast crude mitochondria were incubated with digitonin (0%, 0.01%, 0.02%, 0.04%, 0.06%, 0.08%, and 0.1% [w/v]) and 1% Triton (w/v) in the presence of 300 μg/ml proteinase K (40 μl reaction size, 15 min, RT). Proteinase K was then inhibited with the addition of 7 mM PMSF. Each reaction was precipitated with 25% trichloroacetic acid (20 min, on ice) and resuspended in in 40 μl 1× lithium dodecyl sulfate (LDS) sample buffer containing 4% β-mercaptoethanol (BME). Ten microliters of each reaction was analyzed via western blot as described below.

Schmitz J.M., Wolters J.F., Murray N.H., Guerra R.M., Bingman C.A., Hittinger C.T, & Pagliarini D.J. (2023). Aim18p and Aim46p are chalcone isomerase domain–containing mitochondrial hemoproteins in Saccharomyces cerevisiae. The Journal of Biological Chemistry, 299(3), 102981.

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Publication 2023

2-Mercaptoethanol Biological Assay Buffers Digitonin dodecyl sulfate, lithium salt Endopeptidase K Mitochondria Trichloroacetic Acid Western Blotting Yeast, Dried

Isolation and Analysis of Mitochondrial Complexes

HEK293T (ATCC) cells were maintained in media composed of DMEM (high glucose; Biowest), sodium pyruvate (Thermo Fisher Scientific), stable glutamine (Biowest), non-essential amino acids (Thermo Fisher Scientific), 10% fetal bovine serum (EurX), and penicillin/streptomycin solution (Thermo Fisher Scientific). Cells were cultured to confluence on six 9 cm plates. Mitochondria were isolated by differential fractionation previously described^{42 (link)} with minor adjustments. Briefly, cells were washed 2 × with ice-cold PBS. The PBS buffer was removed and the cells were scraped in ice-cold NKM buffer (1 mM Tris–HCl pH 7.4, 130 mM NaCl, 5 mM KCl, 7.5 mM MgCl) and incubated on ice for 5 min. Then 100 μl of 10xHomB buffer (225 mM mannitol, 75 mM sucrose, 10 mM HEPES–NaOH pH 7.8, 10 mM EDTA) per 1 mL of homogenate with protease and phosphatase inhibitors was added and homogenized using a Dounce homogenizer with nuclei release and integrity monitored microscopically. The nuclear fraction was separated using two rounds of centrifugation at 900 × g for 5 min at 4 °C. The supernatant was then centrifuged at 10,000 × g for 10 min to isolate crude mitochondrial pellet. The pellet was suspended in Blue Native protein extraction buffer, dosed and ATP synthase complex from 400 µg

μ

of protein was extracted with 2% Digitonin. The dimers and monomers were separated in 3–12% gradient gel followed by SDS-PAGE separation in second dimension and bands from 5–20 kDa region from Coomassie stained gel from the monomers and dimers were subjected to mass spectrometric analysis.

Panja C., Wiesyk A., Niedźwiecka K., Baranowska E, & Kucharczyk R. (2023). ATP synthase interactome analysis identifies a new subunit l as a modulator of permeability transition pore in yeast. Scientific Reports, 13, 3839.

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Publication 2023

Amino Acids, Essential Buffers Cell Nucleus Cells Centrifugation Cold Temperature Digitonin Edetic Acid Fetal Bovine Serum Fractionation, Chemical Glucose Glutamine HEPES inhibitors Mannitol Mass Spectrometry Mitochondria Nitric Oxide Synthase Penicillins Peptide Hydrolases Phosphoric Monoester Hydrolases Proteins Pyruvate SDS-PAGE Sodium Sodium Chloride Streptomycin Sucrose Tromethamine

Mitochondrial ATP Synthase Complex Analysis

Two-dimensional gel electrophoresis was based on the protocol of Schamel^{43 (link)} with slight modifications. Briefly, the ATP synthase complexes were liberated from inner mitochondrial membrane of isolated mitochondria by incubation with 1–2% digitonin in extraction buffer (30 mM HEPES, 150 mM potassium acetate, 12% glycerol, 2 mM 6-aminocaproic acid, 1 mM EGTA, protease inhibitor cocktail tablets EDTA-free (Roche), pH 7.4) for different time intervals up to 60 min and separated using NativePAGE™ 3–12% Bis–Tris Gels (Thermo Fisher Scientific) to separate monomeric and dimeric ATP synthase complexes^{44 (link)}. For second dimensional analysis the lanes were cut from the gel and placed in SDS-PAGE running buffer (25 mM Tris, 192 mM Glycine, 0.1% SDS, pH 8.3 with 1% β-mercaptoethanol), heated in a microwave for 10 secs and incubated for another 10 min in a shaker. The gel strips were then loaded on the top of a 16% SDS-PAGE gel, and electrophoresis was conducted under denaturing conditions. Then the gel was stained with Coomassie blue or silver staining and bands cut-off were analyzed by mass spectrometry. For Western blotting proteins from the gel were transferred into PVDF or nitrocellulose membranes using iBlot system (Thermo Fisher Scientific). For SDS-PAGE analysis of steady state level of proteins, yeast cells were disrupted by alkaline lysis with NaOH/TCA^{45 (link)}. Western blot analysis was performed using the polyclonal rabbit anti-Mco10 antibody, anti-ATP synthase subunits antibodies (gifts from Marie-France Giraud, Bordeaux, France and Martin van der Laan, Germany), anti-Rip1 and Cob1 antibodies (provided by dr hab. Ulrike Topf, IBB PAS) or anti-Cox2 (Thermo Fisher Scientific).

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Publication 2023

2-Mercaptoethanol 6-Aminocaproic Acid Anti-Antibodies Antibodies Antibodies, Anti-Idiotypic Bistris Buffers Cells Coomassie blue Digitonin Edetic Acid Egtazic Acid Electrophoresis Electrophoresis, Gel, Two-Dimensional Gifts Glycerin Glycine HEPES Mass Spectrometry Microwaves Mitochondria Mitochondrial Membrane, Inner Nitric Oxide Synthase Nitrocellulose polyvinylidene fluoride Potassium Acetate Protease Inhibitors Proteins Protein Subunits PTGS2 protein, human Rabbits Saccharomyces cerevisiae SDS-PAGE Tissue, Membrane Tromethamine Western Blot

Top products related to «Digitonin»

Digitonin by Merck Group

Sourced in United States, Germany, Italy, Finland, Sao Tome and Principe, Japan

Digitonin is a glycoside extracted from the plant Digitalis purpurea. It is a non-ionic detergent commonly used in laboratory settings to solubilize and permeabilize cell membranes, allowing for the extraction and analysis of cellular components.

Protease inhibitor cocktail by Roche

Sourced in United States, Switzerland, Germany, China, United Kingdom, France, Canada, Japan, Italy, Australia, Austria, Sweden, Spain, Cameroon, India, Macao, Belgium, Israel

Protease inhibitor cocktail is a laboratory reagent used to inhibit the activity of proteases, which are enzymes that break down proteins. It is commonly used in protein extraction and purification procedures to prevent protein degradation.

Oxygraph 2k by Oroboros

Sourced in Austria

The Oxygraph-2k is a high-performance respirometer designed for precise measurement of oxygen consumption and production in biological samples. It provides real-time monitoring of oxygen levels, making it a valuable tool for researchers in the fields of cell biology, physiology, and bioenergetics.

Digitonin by Thermo Fisher Scientific

Sourced in United States

Digitonin is a natural detergent derived from the plant Digitalis purpurea. It is commonly used in biochemical research applications as a tool for cell membrane permeabilization and the extraction of proteins and other biomolecules from cells.

Digitonin by Promega

Sourced in United States

Digitonin is a natural extract from the plant Digitalis purpurea. It is a non-ionic detergent used to permeabilize cell membranes in various laboratory applications.

Minelute pcr purification kit by Qiagen

Sourced in Germany, United States, United Kingdom, Italy, Japan, Netherlands, China, France, Switzerland

The MinElute PCR Purification Kit is a laboratory equipment product designed for the efficient purification of PCR amplicons. It utilizes a silica-membrane-based technology to capture and purify DNA fragments from PCR reactions, allowing for the removal of primers, nucleotides, enzymes, and other impurities.

Lipofectamine 2000 by Thermo Fisher Scientific

Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Italy, Switzerland, Australia, Spain, Belgium, Denmark, Singapore, India, Netherlands, Sweden, New Zealand, Portugal, Poland, Israel, Lithuania, Hong Kong, Argentina, Ireland, Austria, Czechia, Cameroon, Taiwan, Province of China, Morocco

Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.

Protease inhibitor cocktail by Merck Group

Sourced in United States, Germany, China, United Kingdom, Italy, Japan, Sao Tome and Principe, France, Canada, Macao, Switzerland, Spain, Australia, Israel, Hungary, Ireland, Denmark, Brazil, Poland, India, Mexico, Senegal, Netherlands, Singapore

The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.

Nativepage sample buffer by Thermo Fisher Scientific

Sourced in United States, Japan

NativePAGE sample buffer is a product designed for use in Native PAGE (Polyacrylamide Gel Electrophoresis) techniques. It is used to prepare and load protein samples for electrophoretic separation under non-denaturing conditions, preserving the native structure and interactions of proteins.

Bovine serum albumin (bsa) by Merck Group

Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico

Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.

What are the key properties and uses of Digitonin?

Digitonin is a naturally occurring saponin compound derived from the plant Digitalis purpurea. It is commonly used as a detergent and permeabilizing agent in biochemical and cell biology research, particularly for the study of membrane proteins and organellar structures. Digitonin interacts with cholesterol in cell membranes, causing them to become more permeable and allowing for the extraction or fractionation of cellular components. It is a valuable tool for investigating subcellular organization, protein-protein interactions, and the function of membrane-bound proteins.

How can researchers optimize the use of Digitonin in their experiments?

Researchers should carefully optimize Digitonin concentrations and incubation times to preserve cellular integrity and target specificity when using this powerful reagent. The optimal Digitonin concentration can vary depending on the cell type, experimental conditions, and the specific application. Researchers should conduct dose-response experiments to determine the minimum effective concentration that achieves the desired level of permeabilization without compromising cell viability or the integrity of the target structures.

Are there different variations or types of Digitonin, and how do they differ in their applications?

Yes, there are different variations of Digitonin available. While the most common form is extracted from Digitalis purpurea, there are also synthetic or semi-synthetic versions of Digitonin that may have slightly different properties and uses. Researchers should carefully evaluate the specific characteristics of each Digitonin type to ensure they select the most appropriate one for their experimental needs, such as preserving the activity of certain membrane proteins or maintaining the structural integrity of organelles.

How can PubCompare.ai assist researchers in optimizing their Digitonin protocols?

PubCompare.ai's AI-driven reproducibility platform can help researchers optimize their Digitonin protocols in several ways. First, the platform allows you to efficiently screen the protocol literature to identify the most effective Digitonin protocols for your specific research goals. The AI-powered analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy. Secndly, PubCompare.ai's innovative tools can assist you in identifying the critical insights needed to fine-tune your Digitonin protocols, ensuring you achieve the desired level of permeabilization and cellular integrity.

More about "Digitonin"

Digitonin is a naturally occurring saponin compound derived from the Foxglove plant (Digitalis purpurea).
It is widely used as a detergent and permeabilizing agent in biochemical and cell biology research, particularly for the study of membrane proteins and organellar structures.
Digitonin interacts with cholesterol in cell membranes, causing them to become more permeable and allowing for the extraction or fractionation of cellular components.
It is a valuable tool for investigating subcellular organization, protein-protein interactions, and the function of membrane-bound proteins.
Researchers often use Digitonin in conjunction with other reagents like Protease inhibitor cocktails, Oxygraph-2k systems, MinElute PCR Purification Kits, Lipofectamine 2000 transfection reagent, and NativePAGE sample buffer to optimize their experimental protocols.
Bovine serum albumin (BSA) is another common additive used to stabilize proteins and reduce non-specific binding when working with Digitonin.
When using Digitonin, it is important to carefully optimize the concentration and incubation time to preserve cellular integrity and target specificity.
Digitogin can be a powerful but potentially disruptive tool, so researchers must balance its permeabilizing effects with the need to maintain the native structure and function of their samples.
By leveraging the insights gained from the literature and employing innovative AI-driven platforms like PubCommpare.ai, scientists can improve the efficacy and reproducibility of their Digitonin-based research protocols.