Structured Review

Millipore tris
Purification of BmooPAi from B. moojeni snake venom. (a) Separation on DEAE-Sephacel ion-exchange chromatography: crude venom (200 mg) was applied to the column (2.5 × 20 cm) and elution was carried out at a flow rate of 20 mL/h with ammonium bicarbonate (Ambic) buffer gradients, pH 7.8, from 0.05 M to 0.6 M. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (b) Separation on Sephadex G-75 molecular exclusion chromatography: fraction D7 was applied to the column (1.0 × 100 cm) and elution with 0.05 M ammonium bicarbonate was achieved at a flow rate of 20 mL/h. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (c) Separation by affinity chromatography on a HiTrap Heparin HP column using the ÄKTApurifier HPLC system: fraction D7S2 was applied to the column (5 × 1 mL), previously equilibrated with 20 mM <t>Tris-HCl</t> buffer (pH 7.0) containing 5 mM calcium chloride. The samples were eluted with an increasing concentration gradient of 20 mM Tris-HCl buffer (pH 7.0) containing 2.0 M sodium chloride, and the absorbance of the fractions was monitored at 280 nm. Fractions of 1.0 mL/tube were collected at a flow rate of 30 mL/h. (d) <t>SDS-PAGE</t> in 14% (w/v) polyacrylamide, Tris-glycine buffer, pH 8.3, and 20 mA. Lanes: 1, standard proteins; 2, reduced crude venom of B. moojeni ; 3, reduced BmooPAi; 4, nonreduced BmooPAi. The gel was stained with Coomassie blue R-250. (e) Reverse-phase HPLC on a C2C18 column (4.6 × 100 mm) equilibrated with 0.1% trifluoroacetic acid (TFA) and eluted with a linear concentration gradient from 0 to 100% of solution B (70% acetonitrile in 0.1% TFA).
Tris, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 262 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "A New Platelet-Aggregation-Inhibiting Factor Isolated from Bothrops moojeni Snake Venom"

Article Title: A New Platelet-Aggregation-Inhibiting Factor Isolated from Bothrops moojeni Snake Venom

Journal: BioMed Research International

doi: 10.1155/2017/4315832

Purification of BmooPAi from B. moojeni snake venom. (a) Separation on DEAE-Sephacel ion-exchange chromatography: crude venom (200 mg) was applied to the column (2.5 × 20 cm) and elution was carried out at a flow rate of 20 mL/h with ammonium bicarbonate (Ambic) buffer gradients, pH 7.8, from 0.05 M to 0.6 M. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (b) Separation on Sephadex G-75 molecular exclusion chromatography: fraction D7 was applied to the column (1.0 × 100 cm) and elution with 0.05 M ammonium bicarbonate was achieved at a flow rate of 20 mL/h. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (c) Separation by affinity chromatography on a HiTrap Heparin HP column using the ÄKTApurifier HPLC system: fraction D7S2 was applied to the column (5 × 1 mL), previously equilibrated with 20 mM Tris-HCl buffer (pH 7.0) containing 5 mM calcium chloride. The samples were eluted with an increasing concentration gradient of 20 mM Tris-HCl buffer (pH 7.0) containing 2.0 M sodium chloride, and the absorbance of the fractions was monitored at 280 nm. Fractions of 1.0 mL/tube were collected at a flow rate of 30 mL/h. (d) SDS-PAGE in 14% (w/v) polyacrylamide, Tris-glycine buffer, pH 8.3, and 20 mA. Lanes: 1, standard proteins; 2, reduced crude venom of B. moojeni ; 3, reduced BmooPAi; 4, nonreduced BmooPAi. The gel was stained with Coomassie blue R-250. (e) Reverse-phase HPLC on a C2C18 column (4.6 × 100 mm) equilibrated with 0.1% trifluoroacetic acid (TFA) and eluted with a linear concentration gradient from 0 to 100% of solution B (70% acetonitrile in 0.1% TFA).
Figure Legend Snippet: Purification of BmooPAi from B. moojeni snake venom. (a) Separation on DEAE-Sephacel ion-exchange chromatography: crude venom (200 mg) was applied to the column (2.5 × 20 cm) and elution was carried out at a flow rate of 20 mL/h with ammonium bicarbonate (Ambic) buffer gradients, pH 7.8, from 0.05 M to 0.6 M. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (b) Separation on Sephadex G-75 molecular exclusion chromatography: fraction D7 was applied to the column (1.0 × 100 cm) and elution with 0.05 M ammonium bicarbonate was achieved at a flow rate of 20 mL/h. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (c) Separation by affinity chromatography on a HiTrap Heparin HP column using the ÄKTApurifier HPLC system: fraction D7S2 was applied to the column (5 × 1 mL), previously equilibrated with 20 mM Tris-HCl buffer (pH 7.0) containing 5 mM calcium chloride. The samples were eluted with an increasing concentration gradient of 20 mM Tris-HCl buffer (pH 7.0) containing 2.0 M sodium chloride, and the absorbance of the fractions was monitored at 280 nm. Fractions of 1.0 mL/tube were collected at a flow rate of 30 mL/h. (d) SDS-PAGE in 14% (w/v) polyacrylamide, Tris-glycine buffer, pH 8.3, and 20 mA. Lanes: 1, standard proteins; 2, reduced crude venom of B. moojeni ; 3, reduced BmooPAi; 4, nonreduced BmooPAi. The gel was stained with Coomassie blue R-250. (e) Reverse-phase HPLC on a C2C18 column (4.6 × 100 mm) equilibrated with 0.1% trifluoroacetic acid (TFA) and eluted with a linear concentration gradient from 0 to 100% of solution B (70% acetonitrile in 0.1% TFA).

Techniques Used: Purification, Ion Exchange Chromatography, Flow Cytometry, Chromatography, Affinity Chromatography, High Performance Liquid Chromatography, Concentration Assay, SDS Page, Staining

2) Product Images from "Effect of Fengycin, a Lipopeptide Produced by Bacillus subtilis, on Model Biomembranes"

Article Title: Effect of Fengycin, a Lipopeptide Produced by Bacillus subtilis, on Model Biomembranes

Journal:

doi: 10.1529/biophysj.107.114090

Second heating scan of fengycin and/or DPPC vesicles in a 10 mM Tris, 150 mM NaCl buffer at pH 7.4. ( a ) DPPC vesicles 1 mM; ( b ) DPPC/fengycin (300:1), [fengycin] = 3.3 μ M; ( c ) DPPC/fengycin (37.5:1), [fengycin] = 25.8 μ
Figure Legend Snippet: Second heating scan of fengycin and/or DPPC vesicles in a 10 mM Tris, 150 mM NaCl buffer at pH 7.4. ( a ) DPPC vesicles 1 mM; ( b ) DPPC/fengycin (300:1), [fengycin] = 3.3 μ M; ( c ) DPPC/fengycin (37.5:1), [fengycin] = 25.8 μ

Techniques Used:

Surface pressure increase after injection of fengycin under a DPPC monolayer previously compressed at a defined initial surface pressure. Subphase is a 10 mM Tris, 150 mM NaCl at pH 7.4. Temperature is 30°C. Fengycin is injected into the subphase
Figure Legend Snippet: Surface pressure increase after injection of fengycin under a DPPC monolayer previously compressed at a defined initial surface pressure. Subphase is a 10 mM Tris, 150 mM NaCl at pH 7.4. Temperature is 30°C. Fengycin is injected into the subphase

Techniques Used: Injection

3) Product Images from "Tandem Mass Spectrometry Measurement of the Collision Products of Carbamate Anions Derived from CO2 Capture Sorbents: Paving the Way for Accurate Quantitation"

Article Title: Tandem Mass Spectrometry Measurement of the Collision Products of Carbamate Anions Derived from CO2 Capture Sorbents: Paving the Way for Accurate Quantitation

Journal: Journal of the American Society for Mass Spectrometry

doi: 10.1007/s13361-011-0161-5

CO 2 capture amines for which chemistry with bicarbonate (specifically carbamate formation) was investigated. 1 = 2-aminoethanol (ethanolamine, MEA); 2 = 1,2-diaminoethane (ethylenediamine, EN); 3 = piperazine (PZ); 4 = 2-amino-2-methyl-1-propanol (AMP); 5 = 2-amino-2-(hydroxymethyl)propane-1,3-diol ( Tris ); 6 = 2,2'-iminodiethanol (diethanolamine, DEA)
Figure Legend Snippet: CO 2 capture amines for which chemistry with bicarbonate (specifically carbamate formation) was investigated. 1 = 2-aminoethanol (ethanolamine, MEA); 2 = 1,2-diaminoethane (ethylenediamine, EN); 3 = piperazine (PZ); 4 = 2-amino-2-methyl-1-propanol (AMP); 5 = 2-amino-2-(hydroxymethyl)propane-1,3-diol ( Tris ); 6 = 2,2'-iminodiethanol (diethanolamine, DEA)

Techniques Used: Microelectrode Array

Related Articles

Centrifugation:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: After centrifugation at 20,000 rpm for 1 hour at 4 °C, cleared lysate was loaded onto a Ni-NTA FF (GE) affinity column, eluted by an imidazole gradient and the tag removed by overnight incubation at 4 °C with TEV protease. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: After centrifugation at 4 °C for 10 min, the solution is divided into two phases and the middle of the two phases is protein membrane. .. Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA).

Synthesized:

Article Title: Light-induced formation of partially reduced oxygen species limits the lifetime of photosystem 1-based biocathodes
Article Snippet: Synthesis of CuTPA CuTPA (C18 H18 CuN6 O6 , Supplementary Fig. , molecular weight: 477.92 g mol−1 ) was synthesized as follows. .. Tris(2-pyridylmethyl)amine (Sigma-Aldrich, 62.4 mg; 0.2 mmol) was dissolved in absolute EtOH (11 mL).

Immunostaining:

Article Title: Bioinspired neuron-like electronics
Article Snippet: Paragraph title: 2. Tissue clearing and immunostaining. ... If necessary to improve light penetration for microscopy imaging, slices could be further incubated in Tris (Sigma-Aldrich) with 2% SDS (Sigma-Aldrich) and 0.3 M glycine at pH 6.8 at RT for 3 days then re-stained with DAPI.

Electrophoresis:

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503). .. Bead elutions were analyzed by Coomassie Brilliant Blue (SERVA Electrophoresis GmbH, 17,521) staining and anti-His western blotting.

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) ! .. Qiagen Plasmid Plus midi kit (Qiagen, cat. no. 12943) TAE electrophoresis buffer Tris base (Trizma base; Sigma-Aldrich, cat. no. T6066) NaOH

Sandwich ELISA:

Article Title: Effect of oral genistein administration in early and late phases of allergic encephalomyelitis
Article Snippet: For assessing the concentration of cytokines in CNS, 100 mg of frontal and temporal lobes of each mouse was homogenized in 10 ml of an extracting solution containing 50 µM tris (Sigma-Aldrich, USA), 2 mM EDTA (Merck, Germany), 0.1 M NaCl (Sigma-Aldrich, USA), 1 mM dithiotheritol (Merck, Germany), 200 µM phenylmethylsulfonyl fluoride (Merck, Germany), 1 µg/ml chymostatin (Sigma-Aldrich, USA), and 1 µg/ml trypsin inhibitor (Sigma-Aldrich, USA). .. The supernatant was analyzed for IFN-γ, TNF-α, IL-12 (as the proinflammatory cytokines), and IL-10 (as an anti-inflammatory cytokine) concentrations using sandwich-based enzyme-linked immunosorbent assay (ELISA) kits (eBioscience, USA) by following the manufacturer’s instruction.

Incubation:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: After centrifugation at 20,000 rpm for 1 hour at 4 °C, cleared lysate was loaded onto a Ni-NTA FF (GE) affinity column, eluted by an imidazole gradient and the tag removed by overnight incubation at 4 °C with TEV protease. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Briefly, GST-tagged fusion proteins were directly immobilized on glutathione-Sepharose beads (GE Healthcare, GE17–0756–01) from E. coli extracts and incubated with purified proteins for 1 h at 4°C in PBS under rotation. .. Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503).

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma). .. The solution was incubated at 55°C with vigorous orbital shaking using a Thermomixer (Eppendorf).

Article Title: Bioinspired neuron-like electronics
Article Snippet: .. If necessary to improve light penetration for microscopy imaging, slices could be further incubated in Tris (Sigma-Aldrich) with 2% SDS (Sigma-Aldrich) and 0.3 M glycine at pH 6.8 at RT for 3 days then re-stained with DAPI. .. Brain slices were glued at their edge to the bottom of 50-mm-diameter petri dishes by Devcon 5 Minute Epoxy (ITW Polymers Adhesives) and then incubated in RI matching solution PROTOS (diatrizoic acid, Sigma-Aldrich; N-Methyl-D-glucamine, Sigma-Aldrich; OptiPrep, Accurate Chemical and Scientific) or 80% glycerol:20% PBS (glycerol, Sigma-Aldrich) 24 h prior to microscopy imaging.

Expressing:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Paragraph title: 2.1 Protein expression and purification ... Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Bradford Assay:

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA). .. The Bradford assay with Bradford reagents (Thermo Fischer Scientific, Waltham, MA, USA) was supplied to measure the protein concentration of the WAT samples.

Modification:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: The catalytic domain (residues 243–530) of human SHP-1 was subcloned into a modified pET-32 expression vector and expressed in Rosetta (DE3) Escherichia coli . .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Western Blot:

Article Title: Sintokamide A Is a Novel Antagonist of Androgen Receptor That Uniquely Binds Activation Function-1 in Its Amino-terminal Domain *
Article Snippet: Proteins were extracted with lysis buffer and subjected to click chemistry conditions for 3 h at 25 °C in buffer containing 0.1% SDS, 5% t -butyl alcohol, 100 μm tris[(1-benzyl-1H -1,2,3-triazol-4-yl)methyl]amine (Sigma), 1 mm tris(2-carboxyethyl)phosphine, 100 μm biotin-azide reagent, and 1 mm CuSO4 . .. Protein-SINT1 complexes were separated by 10% SDS-PAGE and subjected to Western blotting analysis using AR-N20 (1:1000) and biotin (1:1000) antibodies.

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Paragraph title: Coimmunoprecipitation experiments, zebrafish protein lysate extraction, Coomassie gels and western blot analysis ... Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503).

Chromatography:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Following a second affinity column to remove His-tagged TEV and uncut protein, SHP-1 was further purified by anion exchange chromatography using a Resource Q (GE) column. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Protease Inhibitor:

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: Protein extraction and in-solution digestion The iPSC cell pellets were lysed on ice by using a lysis buffer containing 8 M urea (EuroBio), 40 mM Tris-base (Sigma-Aldrich), 10 mM DTT (AppliChem) and complete protease inhibitor cocktail (Roche). .. The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich).

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: One mL lysis buffer (protein extraction kit, P1250-50, Applygen, Beijing) and one μL protease inhibitor cocktail (50x, P1265-0.5, Applygen, Beijing) (1:1,000) were added to the tissue. .. Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA).

Generated:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP). .. SHP-1 variants (C453S, Q252A, Q252R, C453S/Q252A, and C453S/Q252R) were generated using the QuikChange Lightning Site-Directed Mutagenesis Kit (Stratagene).

Article Title: Tandem Mass Spectrometry Measurement of the Collision Products of Carbamate Anions Derived from CO2 Capture Sorbents: Paving the Way for Accurate Quantitation
Article Snippet: Reagents and Carbamate Synthesis The carbamate derivatives of capture amines were generated by mixing equimolar amounts (1–2 mmol) of the respective amine and NaHCO3 (Sigma, Sydney, Australia, 99.9%) in 15 mL charcoal-filtered water (R > 18 MΩ). .. Reagent purity: MEA (Sigma, Sydney, Australia, > 99%), EN (Sigma, Sydney, Australia, 99%), PZ (Sigma, Sydney, Australia, 99 %), AMP (Fluka, Sydney, Australia, 99%), DEA (Aldrich, Sydney, Australia, > 99.5%), Tris (Sigma, Sydney, Australia, > 99%).

Cytokine Assay:

Article Title: Effect of oral genistein administration in early and late phases of allergic encephalomyelitis
Article Snippet: Paragraph title: Brain cytokine assay ... For assessing the concentration of cytokines in CNS, 100 mg of frontal and temporal lobes of each mouse was homogenized in 10 ml of an extracting solution containing 50 µM tris (Sigma-Aldrich, USA), 2 mM EDTA (Merck, Germany), 0.1 M NaCl (Sigma-Aldrich, USA), 1 mM dithiotheritol (Merck, Germany), 200 µM phenylmethylsulfonyl fluoride (Merck, Germany), 1 µg/ml chymostatin (Sigma-Aldrich, USA), and 1 µg/ml trypsin inhibitor (Sigma-Aldrich, USA).

Imaging:

Article Title: Bioinspired neuron-like electronics
Article Snippet: .. If necessary to improve light penetration for microscopy imaging, slices could be further incubated in Tris (Sigma-Aldrich) with 2% SDS (Sigma-Aldrich) and 0.3 M glycine at pH 6.8 at RT for 3 days then re-stained with DAPI. .. Brain slices were glued at their edge to the bottom of 50-mm-diameter petri dishes by Devcon 5 Minute Epoxy (ITW Polymers Adhesives) and then incubated in RI matching solution PROTOS (diatrizoic acid, Sigma-Aldrich; N-Methyl-D-glucamine, Sigma-Aldrich; OptiPrep, Accurate Chemical and Scientific) or 80% glycerol:20% PBS (glycerol, Sigma-Aldrich) 24 h prior to microscopy imaging.

Protein Concentration:

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: Supernatants were collected and protein concentration was measured by DC Protein Assay kit® (Bio-Rad). .. The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma).

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA). .. The Bradford assay with Bradford reagents (Thermo Fischer Scientific, Waltham, MA, USA) was supplied to measure the protein concentration of the WAT samples.

Sequencing:

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich). .. The samples were then redissolved by 100 mM NH4 HCO3 and were digested with sequencing-grade porcine trypsin (Promega) at a protease/protein ratio of 1:40 overnight at 37 °C [ ].

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: MO(s) (Gene Tools, must be custom designed to target sequence) Agarose (ISC BioExpress, cat. no. E-3119-500) ASW (Instant Ocean, cat. no. SS15-10) Dextran, Alexa Fluor 555 dye (20 mg ml−1 in nuclease-free water; Invitrogen, cat. no. ) Dextran, Alexa Fluor 488 dye (20 mg ml−1 in nuclease-free water; Invitrogen, cat. no. ) EDTA (Sigma-Aldrich, cat. no. E5134-500G) Ethanol (Pharmco-Aaper, cat. no. 111000200) Glacial acetic acid (Fisher Scientific, cat. no. BP2401SI-212) Glycogen (20 mg ml−1 ; Roche Applied Science, cat. no. 10901393001) Isopropanol (Fisher Scientifc, cat. no. A4164) ! .. Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) !

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma). .. Protein alkylation was done by adding 10 mM iodoacetamide (Sigma) and incubating with vigorous shaking in the dark for 20 min. To digest the proteins, we added in the following order: 150 μl of 50 mM NH4 HCO3 , 2.5 μl of 1% ProteaseMAX dissolved in 50 mM NH4 HCO3 and 1:100 (enzyme/protein, w/w) sequencing grade trypsin (Promega) to a final reaction volume of 500 ml.

Sonication:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Cell pellets were resuspended in lysis buffer (20 mM Tris pH 8.5, 200 mM NaCl, 1.5 mg/mL lysozyme) supplemented with protease inhibitors, and lysed by repetitive freeze/thaw cycles and sonication. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: The resulted mixture were sonicated in 4 °C for 5 mins using a VialTweeter device (Hielscher-Ultrasound Technology) and centrifuged at 21,130 g and 4 °C for 1 hr to remove the insoluble material. .. The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich).

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: Protein pellets were washed once in methanol, air-dried for 10 min and resuspended the in 200 μl of a buffer containing 4 M urea, 50 mM NH4 HCO3 and 0.1% ProteaseMax surfactant (Promega) with a brief sonication pulse using a Sonic Dismembrator Model 100 (Fisher Scientific). .. The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma).

Injection:

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: Paragraph title: For morpholino injection ... Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) !

Binding Assay:

Article Title: Sintokamide A Is a Novel Antagonist of Androgen Receptor That Uniquely Binds Activation Function-1 in Its Amino-terminal Domain *
Article Snippet: Paragraph title: Binding Assay of SINT1 to Full-length AR ... Proteins were extracted with lysis buffer and subjected to click chemistry conditions for 3 h at 25 °C in buffer containing 0.1% SDS, 5% t -butyl alcohol, 100 μm tris[(1-benzyl-1H -1,2,3-triazol-4-yl)methyl]amine (Sigma), 1 mm tris(2-carboxyethyl)phosphine, 100 μm biotin-azide reagent, and 1 mm CuSO4 .

Molecular Weight:

Article Title: Light-induced formation of partially reduced oxygen species limits the lifetime of photosystem 1-based biocathodes
Article Snippet: Synthesis of CuTPA CuTPA (C18 H18 CuN6 O6 , Supplementary Fig. , molecular weight: 477.92 g mol−1 ) was synthesized as follows. .. Tris(2-pyridylmethyl)amine (Sigma-Aldrich, 62.4 mg; 0.2 mmol) was dissolved in absolute EtOH (11 mL).

DC Protein Assay:

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: Supernatants were collected and protein concentration was measured by DC Protein Assay kit® (Bio-Rad). .. The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma).

Mutagenesis:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP). .. SHP-1 variants (C453S, Q252A, Q252R, C453S/Q252A, and C453S/Q252R) were generated using the QuikChange Lightning Site-Directed Mutagenesis Kit (Stratagene).

Microelectrode Array:

Article Title: Tandem Mass Spectrometry Measurement of the Collision Products of Carbamate Anions Derived from CO2 Capture Sorbents: Paving the Way for Accurate Quantitation
Article Snippet: .. Reagent purity: MEA (Sigma, Sydney, Australia, > 99%), EN (Sigma, Sydney, Australia, 99%), PZ (Sigma, Sydney, Australia, 99 %), AMP (Fluka, Sydney, Australia, 99%), DEA (Aldrich, Sydney, Australia, > 99.5%), Tris (Sigma, Sydney, Australia, > 99%). ..

Reflux:

Article Title: Light-induced formation of partially reduced oxygen species limits the lifetime of photosystem 1-based biocathodes
Article Snippet: Tris(2-pyridylmethyl)amine (Sigma-Aldrich, 62.4 mg; 0.2 mmol) was dissolved in absolute EtOH (11 mL). .. Cu(NO3 )2 ·3H2 O (25.4 mg; 0.1 mmol, 0.5 eq.) was added and the mixture was stirred at reflux temperature for 30 min. EtOH was later removed by rotary evaporation.

Labeling:

Article Title: The Retinal Ganglion Cell Transportome Identifies Proteins Transported to Axons and Presynaptic Compartments in the Visual System In Vivo
Article Snippet: Paragraph title: Sample processing for direct identification of biotin labeled proteins using DiDBiT ... The protein suspension was reduced by adding of 5 mM Tris(2-carboxyethyl)phosphine (Sigma).

Article Title: Single cell-laden protease-sensitive microniches for long-term culture in 3D
Article Snippet: To make TG-PEG precursor solution, Gln-PEG and Lys-MMP-PEG are mixed stoichiometrically in 50 mM Tris (Sigma-Aldrich, cat. no. T5941) pH 7.6 to a final concentration of 30% (v/w). .. The peptides Ac-FKGG-RGDSPG-NH2 (Lys-RGD) and Ac-FKGG-K-FITC (Lys-FITC) are introduced as adhesion ligand and for FITC labeling of the matrix, respectively.

Purification:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP). .. 6×His-tagged SHP-1 was purified over a Ni-NTA FF affinity column, followed by anion exchange chromatography using a Resource Q column and was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM TCEP.

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich). .. Digests were purified with Vydac C18 Silica MicroSpin columns (The Nest Group Inc.).

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Briefly, GST-tagged fusion proteins were directly immobilized on glutathione-Sepharose beads (GE Healthcare, GE17–0756–01) from E. coli extracts and incubated with purified proteins for 1 h at 4°C in PBS under rotation. .. Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503).

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: L-Cysteine (Sigma-Aldrich, cat. no. C7353-100G) MegaClear RNA purification columns (MegaClear kit; Ambion, cat. no. AM1908) mMessage RNA in vitro translation kit (SP6, T7 or T3; e.g., mMessage machine T3; Ambion, cat. no. AM1348) Nuclease-free water (Ambion, cat. no. AM9937) ▴ CRITICAL Use non-DEPC-treated water because DEPC interferes with MO efficiency NucleoBond PC100 (Clontech, cat. no. 740573) Phenol:chloroform:isoamyl alcohol (25:24:1, pH 6.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) ! .. Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) !

Protein Extraction:

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: Paragraph title: Protein extraction and in-solution digestion ... The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich).

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503). .. Protein extraction of embryonic zebrafish and immunoblotting was carried out as described previously [ ].

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: Two mL protein extraction regents were added to the tissue and kept in 4 °C for 40 min to separate lipid completely. .. Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA).

Blocking Assay:

Article Title: Effect of Fengycin, a Lipopeptide Produced by Bacillus subtilis, on Model Biomembranes
Article Snippet: The dry lipid film was hydrated for 1 h at 50°C in a 10 mM Tris (Sigma), 150 mM NaCl (Merck) buffer at pH 7.4. .. The extruder used was equipped with a heating block, which was set to a temperature between 50°C and 55°C, i.e., significantly above the chain-melting transition temperature of the DPPC.

Lysis:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Cell pellets were resuspended in lysis buffer (20 mM Tris pH 8.5, 200 mM NaCl, 1.5 mg/mL lysozyme) supplemented with protease inhibitors, and lysed by repetitive freeze/thaw cycles and sonication. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Article Title: Sintokamide A Is a Novel Antagonist of Androgen Receptor That Uniquely Binds Activation Function-1 in Its Amino-terminal Domain *
Article Snippet: .. Proteins were extracted with lysis buffer and subjected to click chemistry conditions for 3 h at 25 °C in buffer containing 0.1% SDS, 5% t -butyl alcohol, 100 μm tris[(1-benzyl-1H -1,2,3-triazol-4-yl)methyl]amine (Sigma), 1 mm tris(2-carboxyethyl)phosphine, 100 μm biotin-azide reagent, and 1 mm CuSO4 . ..

Article Title: TRIC: an automated alignment strategy for reproducible protein quantification in targeted proteomics
Article Snippet: Protein extraction and in-solution digestion The iPSC cell pellets were lysed on ice by using a lysis buffer containing 8 M urea (EuroBio), 40 mM Tris-base (Sigma-Aldrich), 10 mM DTT (AppliChem) and complete protease inhibitor cocktail (Roche). .. The protein mixtures were reduced by 5 mM tris(carboxyethyl)phosphine (Sigma-Aldrich) and alkylated by 30 mM iodoacetamide (Sigma-Aldrich).

Article Title: Comprehensive map and functional annotation of the mouse white adipose tissue proteome
Article Snippet: And the tissue was homogenized using a spinning blade tissue homogenizer (IKA R104, Janke & Kunkel KG.IKA-werk, Staufen, Germany) on ice for complete lysis. .. Protein dissolution was performed by adding a buffer solution containing seven M urea, two M thiourea, 65 mM DTE, and 83 mM Tris (Sigma-Aldrich, St. Louis, MO, USA).

Mouse Assay:

Article Title: Effect of oral genistein administration in early and late phases of allergic encephalomyelitis
Article Snippet: Brain cytokine assay One day after the last treatment, the mice were sacrificed and their brains were removed. .. For assessing the concentration of cytokines in CNS, 100 mg of frontal and temporal lobes of each mouse was homogenized in 10 ml of an extracting solution containing 50 µM tris (Sigma-Aldrich, USA), 2 mM EDTA (Merck, Germany), 0.1 M NaCl (Sigma-Aldrich, USA), 1 mM dithiotheritol (Merck, Germany), 200 µM phenylmethylsulfonyl fluoride (Merck, Germany), 1 µg/ml chymostatin (Sigma-Aldrich, USA), and 1 µg/ml trypsin inhibitor (Sigma-Aldrich, USA).

SDS Page:

Article Title: Sintokamide A Is a Novel Antagonist of Androgen Receptor That Uniquely Binds Activation Function-1 in Its Amino-terminal Domain *
Article Snippet: Proteins were extracted with lysis buffer and subjected to click chemistry conditions for 3 h at 25 °C in buffer containing 0.1% SDS, 5% t -butyl alcohol, 100 μm tris[(1-benzyl-1H -1,2,3-triazol-4-yl)methyl]amine (Sigma), 1 mm tris(2-carboxyethyl)phosphine, 100 μm biotin-azide reagent, and 1 mm CuSO4 . .. Protein-SINT1 complexes were separated by 10% SDS-PAGE and subjected to Western blotting analysis using AR-N20 (1:1000) and biotin (1:1000) antibodies.

Plasmid Preparation:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: The catalytic domain (residues 243–530) of human SHP-1 was subcloned into a modified pET-32 expression vector and expressed in Rosetta (DE3) Escherichia coli . .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) ! .. Qiagen Plasmid Plus midi kit (Qiagen, cat. no. 12943) TAE electrophoresis buffer Tris base (Trizma base; Sigma-Aldrich, cat. no. T6066) NaOH

Microscopy:

Article Title: Bioinspired neuron-like electronics
Article Snippet: .. If necessary to improve light penetration for microscopy imaging, slices could be further incubated in Tris (Sigma-Aldrich) with 2% SDS (Sigma-Aldrich) and 0.3 M glycine at pH 6.8 at RT for 3 days then re-stained with DAPI. .. Brain slices were glued at their edge to the bottom of 50-mm-diameter petri dishes by Devcon 5 Minute Epoxy (ITW Polymers Adhesives) and then incubated in RI matching solution PROTOS (diatrizoic acid, Sigma-Aldrich; N-Methyl-D-glucamine, Sigma-Aldrich; OptiPrep, Accurate Chemical and Scientific) or 80% glycerol:20% PBS (glycerol, Sigma-Aldrich) 24 h prior to microscopy imaging.

Positron Emission Tomography:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: The catalytic domain (residues 243–530) of human SHP-1 was subcloned into a modified pET-32 expression vector and expressed in Rosetta (DE3) Escherichia coli . .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

In Vitro:

Article Title: Microinjection of mRNA or morpholinos for reverse genetic analysis in the starlet sea anemone, Nematostella vectensis
Article Snippet: L-Cysteine (Sigma-Aldrich, cat. no. C7353-100G) MegaClear RNA purification columns (MegaClear kit; Ambion, cat. no. AM1908) mMessage RNA in vitro translation kit (SP6, T7 or T3; e.g., mMessage machine T3; Ambion, cat. no. AM1348) Nuclease-free water (Ambion, cat. no. AM9937) ▴ CRITICAL Use non-DEPC-treated water because DEPC interferes with MO efficiency NucleoBond PC100 (Clontech, cat. no. 740573) Phenol:chloroform:isoamyl alcohol (25:24:1, pH 6.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) ! .. Phenol:chloroform:isoamyl alcohol (25:24:1, pH 8.0) saturated with 10 mM Tris (Sigma-Aldrich, cat. no. P2069-400ML) !

Spectrophotometry:

Article Title: Forcing the reversibility of a mechanochemical reaction
Article Snippet: UV spectrophotometric measurements for pK a determination Determination of the sulfur pK a values for the studied thiols was conducted through absorbance measurements using a JENWAY 6305 spectrophotometer at λ = 240 nm . .. Determination of the pK a values for L-cysteine, NAC, and 1-mercapto-2-propanol was performed with the addition of 1 mM of tris(2-carboxyethyl)phosphine (Sigma-Aldrich) to prevent dimerization.

Evaporation:

Article Title: Light-induced formation of partially reduced oxygen species limits the lifetime of photosystem 1-based biocathodes
Article Snippet: Tris(2-pyridylmethyl)amine (Sigma-Aldrich, 62.4 mg; 0.2 mmol) was dissolved in absolute EtOH (11 mL). .. Cu(NO3 )2 ·3H2 O (25.4 mg; 0.1 mmol, 0.5 eq.) was added and the mixture was stirred at reflux temperature for 30 min. EtOH was later removed by rotary evaporation.

Concentration Assay:

Article Title: Effect of oral genistein administration in early and late phases of allergic encephalomyelitis
Article Snippet: .. For assessing the concentration of cytokines in CNS, 100 mg of frontal and temporal lobes of each mouse was homogenized in 10 ml of an extracting solution containing 50 µM tris (Sigma-Aldrich, USA), 2 mM EDTA (Merck, Germany), 0.1 M NaCl (Sigma-Aldrich, USA), 1 mM dithiotheritol (Merck, Germany), 200 µM phenylmethylsulfonyl fluoride (Merck, Germany), 1 µg/ml chymostatin (Sigma-Aldrich, USA), and 1 µg/ml trypsin inhibitor (Sigma-Aldrich, USA). ..

Article Title: Forcing the reversibility of a mechanochemical reaction
Article Snippet: Solutions were prepared at a final concentration of 250 μM of the tested thiols in PBS buffer. .. Determination of the pK a values for L-cysteine, NAC, and 1-mercapto-2-propanol was performed with the addition of 1 mM of tris(2-carboxyethyl)phosphine (Sigma-Aldrich) to prevent dimerization.

Article Title: Single cell-laden protease-sensitive microniches for long-term culture in 3D
Article Snippet: .. To make TG-PEG precursor solution, Gln-PEG and Lys-MMP-PEG are mixed stoichiometrically in 50 mM Tris (Sigma-Aldrich, cat. no. T5941) pH 7.6 to a final concentration of 30% (v/w). .. The peptides Ac-FKGG-RGDSPG-NH2 (Lys-RGD) and Ac-FKGG-K-FITC (Lys-FITC) are introduced as adhesion ligand and for FITC labeling of the matrix, respectively.

Staining:

Article Title: Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo
Article Snippet: Afterwards, the beads were washed 3 times with phosphate-buffered saline (PBS; Gibco, 10,010,023) and eluted with 20 mM reduced glutathione (GE Healthcare, GE17–5132–01) in 50 mM Tris (Sigma Aldrich, T1503). .. Bead elutions were analyzed by Coomassie Brilliant Blue (SERVA Electrophoresis GmbH, 17,521) staining and anti-His western blotting.

Article Title: Bioinspired neuron-like electronics
Article Snippet: Slices were placed in fresh PBST containing 1 μg/ml DAPI (Sigma-Aldrich) at RT to let excess antibody diffuse out of the tissue and simultaneously stain nuclear DNA. .. If necessary to improve light penetration for microscopy imaging, slices could be further incubated in Tris (Sigma-Aldrich) with 2% SDS (Sigma-Aldrich) and 0.3 M glycine at pH 6.8 at RT for 3 days then re-stained with DAPI.

Affinity Column:

Article Title: Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition
Article Snippet: Following a second affinity column to remove His-tagged TEV and uncut protein, SHP-1 was further purified by anion exchange chromatography using a Resource Q (GE) column. .. Purified SHP-1 catalytic domain was buffer exchanged to 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (Sigma) (TCEP).

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