ethylenediaminetetraacetic acid edta  (Millipore)


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    Structured Review

    Millipore ethylenediaminetetraacetic acid edta
    Ethylenediaminetetraacetic Acid Edta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ethylenediaminetetraacetic acid edta/product/Millipore
    Average 99 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    ethylenediaminetetraacetic acid edta - by Bioz Stars, 2020-03
    99/100 stars

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    MTT Assay:

    Article Title: Metal-organic Framework Encapsulation Preserves the Bioactivity of Protein Therapeutics
    Article Snippet: .. 2-methylimidazole, zinc acetate dihydrate, ethylenediaminetetraacetic acid (EDTA), ethyl acetate, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT), dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich. .. Human recombinant insulin and insulin sandwich ELISA kit (detection range, 15.60–1,000 pmol/L) were purchased from R & D systems.

    Article Title: Effect of sub-toxic chlorpyrifos on redox sensitive kinases and insulin signaling in rat L6 myotubes
    Article Snippet: 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT), Trypsin EDTA solution were purchased from Hi-media (Mumbai, India). .. Dimethyl sulfoxide (DMSO), insulin, protease-phosphatase inhibitor cocktail (PPI), radioimmunoprecipitation assay (RIPA) buffer with ethylenediaminetetraacetic acid (EDTA) and ethylene glycol tetraacetic acid were purchased from Sigma Aldirch (Missouri, USA).

    Recombinant:

    Article Title: Metal-organic Framework Encapsulation Preserves the Bioactivity of Protein Therapeutics
    Article Snippet: 2-methylimidazole, zinc acetate dihydrate, ethylenediaminetetraacetic acid (EDTA), ethyl acetate, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT), dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich. .. Human recombinant insulin and insulin sandwich ELISA kit (detection range, 15.60–1,000 pmol/L) were purchased from R & D systems.

    Article Title: Development of an LC-MS/MS peptide mapping protocol for the NISTmAb
    Article Snippet: .. Guanidine HCl (Cat #RDD001), Tris(hydroxymethyl)aminomethane (Cat #T6066), Tris(hydroxymethyl)aminomethane HCl (Cat #T5941), ethylenediaminetetraacetic acid (EDTA) (Cat #39692), urea (Cat #U0631), acetic acid (Cat #695084), iodoacetamide (IAM) (Cat #A3221), recombinant porcine trypsin expressed in P. pastoris (Cat #03708985001), trypsin purified from bovine pancreas (Cat #TRYPSEQM-RO) and trypsin purified from porcine pancreas (Cat #T6567) were purchased from Sigma Aldrich. .. Recombinant bovine trypsin expressed in corn (Cat #PRO-313) and recombinant human-2 trypsin expressed in E. coli (Cat # PRO-770) were purchased from ProSpec.

    In Vitro:

    Article Title: Development and Characterization of Nanoparticles for the Delivery of Gemcitabine Hydrochloride
    Article Snippet: .. Appropriate culture medium, Sodium bicarbonate (Sigma, cat. number S5761), 10 mM minimal essential medium (MEM), nonessential aminoacid solution (In vitro gen, cat. number 11140), 100 mM sodium pyruvate (Hyclone, cat. number SH30239), FBS (PAA Laboratories, cat. number A11-043), 10 mg mL−1 bovine insulin in 25 mM HEPES, pH 8.2 (Sigma, cat. number I0516), 2.5% (wt/vol) trypsin solution (Invitrogen, cat. number 15090), 0.5% (wt/vol) phenol red solution (Sigma, cat. number P0290), 0.48 mM versene-EDTA, 0.4% (wt/vol) trypan blue in 0.81% (wt/vol) NaCl, and 0.61% (wt/vol) KH2 PO4 (Sigma, cat. number T8154) were also used. .. Reagents Dimethyl sulfoxide (DMSO; Sigma (cat. number D4540)), Positive control: doxorubicin (Sigma, cat. number D1515), 10% (w/v) TCA, 1% (v/v) acetic acid, 0.057% (w/v) SRB (Fluka, cat. number 86183) in 1% (v/v) acetic acid, and 10 mm unbuffered Tris base solution were used.

    Article Title: Nitric Oxide Releasing Vascular Catheters for Eradicating Bacterial Infection
    Article Snippet: Potassium phosphate monobasic, sodium phosphate dibasic, potassium chloride and sodium chloride, N-acetyl-D-penicillamine (NAP), sodium nitrite, tetrahydrofuran (THF), Ethylenediaminetetraacetic acid (EDTA), hydrochloric acid, sulfuric acid, fibrinogen from bovine plasma, and N, N -dimethylacetamide (DMAc) were obtained from Sigma-Aldrich (St. Louis, MO). .. Autoclaved phosphate buffered saline (PBS), pH 7.4, containing 138 mM NaCl, 2.7 mM KCl, and 10 mM sodium phosphate was used for all in vitro experiments.

    Radio Immunoprecipitation:

    Article Title: Effect of sub-toxic chlorpyrifos on redox sensitive kinases and insulin signaling in rat L6 myotubes
    Article Snippet: .. Dimethyl sulfoxide (DMSO), insulin, protease-phosphatase inhibitor cocktail (PPI), radioimmunoprecipitation assay (RIPA) buffer with ethylenediaminetetraacetic acid (EDTA) and ethylene glycol tetraacetic acid were purchased from Sigma Aldirch (Missouri, USA). ..

    Synthesized:

    Article Title: Highly Sensitive Genosensing Coupling Rolling Circle Amplification with Multiple DNAzyme Cores for DNA Walking.
    Article Snippet: Herein, a highly sensitive electrochemical genosensor is proposed by the construction of an innovative DNA walking machine. .. Herein, a highly sensitive electrochemical genosensor is proposed by the construction of an innovative DNA walking machine.

    Expressing:

    Article Title: Single cell polarity in liquid phase facilitates tumour metastasis
    Article Snippet: .. SkMel2 cells expressing ezrin-GFP were detached using Versene/EDTA, incubated in DMEM for 30 min, pelleted, resuspended in 20% BSA (Sigma 20 µl/10 cm dish) and cryo-immobilised using a high-pressure freezing machine (HPM 010, BAL-TEC). ..

    Microscopy:

    Article Title: A Biomimetic Approach for the Creation of Two-Dimensional Microscale Surface Patterns: Creation of Isolated Immunological Synapses
    Article Snippet: Materials Amine-functionalized microscope slides were purchased from VWR (cat. no. 16001-008) and were cut to 1 in2 pieces using a diamond scribe. .. Glutaraldehyde, hydroxylamine, dimethyl sulfoxide (DMSO), phosphate buffered saline (PBS), ethylenedinitrilotetraacetic acid (EDTA), sodium cyanoborohydride, the fluorescein-5-isothiocyanate (FITC) conjugate of bovine serum albumin (BSA-FITC), and mouse IgA were purchased from Sigma-Alrich; N -succinimidyl-S-acetylthioacetate (SATP), N -(6-(biotinamido)hexyl)-3′-(2′-pyridyldithio)-propionamide (biotin-HPDP), sulfosuccinimidyl acetate (NHS-acetate), N -hydroxysulfosuccinimide (NHS), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) were purchased from Pierce Biotechnology; amine-terminated poly(ethylene glycol) (NH2 -PEG; 10 kDa) and aldehyde-terminated PEG (CHO-PEG; 10 kDa) were purchased from Polysciences, Inc; FITC, AlexaFluor 568 hydrazide sodium salt (AlexaFluor 568), AlexaFluor 568 C5 maleimide (AlexaFluor 568 maleimide), streptavidin, and atto565-biotin were purchased from Invitrogen.

    Mouse Assay:

    Article Title: Carnosine protects cardiac myocytes against lipid peroxidation products
    Article Snippet: Hydrochloric acid gas, 1,1’-carbonyldiimidazole, methyl iodide, 3-(dimethylamino) propanoic acid hydrochloride, ethylenediaminetetraacetic acid (EDTA), ferrozine, EDC (N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide), and N-methylmorpholine, carnosine, were purchased from Sigma-Aldrich and tyrosine-histidine (internal standard: IS) was purchased from Bachem. .. Adult male C57BL/6 mice were obtained from The Jackson Laboratory (Bar Harbor, ME).

    Produced:

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: .. Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO). .. N-succinimidyl S-acetylthioacetate (SATA) (Cat. PI26102), potassium hydroxide pellets, concentrated H2 SO4 (95.0 to 98.0 w/w %), and concentrated HNO3 (70% w/w %) were obtained from Fisher Scientific (Pittsburgh, PA.).

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: .. Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO). .. N-succinimidyl S-acetylthioacetate (SATA) (Cat. PI26102), potassium hydroxide pellets, concentrated H2 SO4 (95.0 to 98.0 w/w %), and concentrated HNO3 (70% w/w %) were obtained from Fisher Scientific (Pittsburgh, PA.).

    Sandwich ELISA:

    Article Title: Metal-organic Framework Encapsulation Preserves the Bioactivity of Protein Therapeutics
    Article Snippet: 2-methylimidazole, zinc acetate dihydrate, ethylenediaminetetraacetic acid (EDTA), ethyl acetate, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT), dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich. .. Human recombinant insulin and insulin sandwich ELISA kit (detection range, 15.60–1,000 pmol/L) were purchased from R & D systems.

    Incubation:

    Article Title: Single cell polarity in liquid phase facilitates tumour metastasis
    Article Snippet: .. SkMel2 cells expressing ezrin-GFP were detached using Versene/EDTA, incubated in DMEM for 30 min, pelleted, resuspended in 20% BSA (Sigma 20 µl/10 cm dish) and cryo-immobilised using a high-pressure freezing machine (HPM 010, BAL-TEC). ..

    other:

    Article Title: Bioconjugated, Single-Use Biosensor for the Detection of Biomarkers of Prostate Cancer
    Article Snippet: Ethylenediaminetetraacetic acid (EDTA) (Cat. EDS) and hydroxylamine (Cat. #255580) were obtained from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: Biochemical characterization of a native group III trypsin ZT from Atlantic cod (Gadus morhua).
    Article Snippet: Chemicals Dimethyl sulfoxide (DMSO), ethylenediaminetetraacetic acid (EDTA), Bis-Tris, glycine, sodium dodecyl sulphate (SDS), dithiothreitol (DTT), benzamidine, 2-mercaptoethanol, phenyl methyl sulphonyl fluoride (PMSF), soybean trypsin inhibitor, tosyl-lysyl-chloromethyl ketone (TLCK), N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK), 2-Amino-2-(hydroxymethyl)-1,3-propanediol (TRIS) and bovine albumin protein standard were from Sigma Aldrich, St. Louis, MO, USA.

    Mass Spectrometry:

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: Human TDP43 peptide (Cat. ab41970) was purchased from Abcam (Cambridge, MS). .. Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO).

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: Human TDP43 peptide (Cat. ab41970) was purchased from Abcam (Cambridge, MS). .. Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO).

    BIA-KA:

    Article Title: Metal-organic Framework Encapsulation Preserves the Bioactivity of Protein Therapeutics
    Article Snippet: 2-methylimidazole, zinc acetate dihydrate, ethylenediaminetetraacetic acid (EDTA), ethyl acetate, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT), dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich. .. The Pierce bicinchoninic acid (BCA) protein assay kit and trypsin were obtained from Thermo Fisher Scientific.

    Article Title: Effect of sub-toxic chlorpyrifos on redox sensitive kinases and insulin signaling in rat L6 myotubes
    Article Snippet: Dimethyl sulfoxide (DMSO), insulin, protease-phosphatase inhibitor cocktail (PPI), radioimmunoprecipitation assay (RIPA) buffer with ethylenediaminetetraacetic acid (EDTA) and ethylene glycol tetraacetic acid were purchased from Sigma Aldirch (Missouri, USA). .. Bicinchoninic Acid (BCA) based protein assay kit was purchased from Thermo Scientific Pierce (Masschussetts, USA).

    Modification:

    Article Title: Metal-organic Framework Encapsulation Preserves the Bioactivity of Protein Therapeutics
    Article Snippet: 2-methylimidazole, zinc acetate dihydrate, ethylenediaminetetraacetic acid (EDTA), ethyl acetate, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT), dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich. .. Dulbecco’s modified eagle medium and Dulbecco’s phosphate-buffered saline (DPBS) were purchased from Gibco.

    Article Title: Pharmacological characterization of the seven human NOX isoforms and their inhibitors
    Article Snippet: 2.1 Reagents Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F-12), Dulbecco׳s modified Eagle medium with 4.5 g/L glucose (DMEM), Roswell Park Memorial Institute (RPMI) 1640 with Glutamax, fetal bovine serum (FBS), penicillin, streptomycin, Dulbecco's Phosphate-Buffered Saline (DPBS), Hanks׳ buffered salt solution with CaCl2 , MgCl2 , glucose and sodium bicarbonate (HBSS), trypsin-EDTA (0.05%), Amplex® Red and Calcein-AM were purchased from Invitrogen. (Dextran T500) was purchased from Pharmacia Fine chemicals Uppsala, Sweden. .. Horseradish peroxidase (HRP), flavin adenine dinucleotide disodium salt hydrate (FAD-Na2 ), phorbol 12-myristate 13-acetate (PMA), ionomycin calcium, superoxide dismutase (SOD), catalase, purified myeloperoxidase, dimethyl sulfoxide (DMSO), hydrogen peroxide 35% (H2 O2 ), 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), ethylenedinitrilotetraacetic acid (EDTA), N,N-bis(carboxymethyl)glycine (NTA), 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N'-tetraacetic acid (BAPTA), 3-sn -phosphatidic acid sodium salt, tetramethyl benzidine, dimethylformamide, MgCl2 , VAS2870 (CAS 722456-31-7), ebselen (CAS 60940-34-3), diapocynin (CAS 29799-22-2) and apocynin (CAS 498-02-2) were purchased from Sigma-Aldrich.

    Article Title: Effect of sub-toxic chlorpyrifos on redox sensitive kinases and insulin signaling in rat L6 myotubes
    Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin were purchased from Gibco Invitrogen (Barcelona, Spain). .. Dimethyl sulfoxide (DMSO), insulin, protease-phosphatase inhibitor cocktail (PPI), radioimmunoprecipitation assay (RIPA) buffer with ethylenediaminetetraacetic acid (EDTA) and ethylene glycol tetraacetic acid were purchased from Sigma Aldirch (Missouri, USA).

    Planar Chromatography:

    Article Title: Nitric Oxide Releasing Vascular Catheters for Eradicating Bacterial Infection
    Article Snippet: Elast-eon E2As was obtained from AorTech International, plc (Scoresby, Victoria, Australia). .. Potassium phosphate monobasic, sodium phosphate dibasic, potassium chloride and sodium chloride, N-acetyl-D-penicillamine (NAP), sodium nitrite, tetrahydrofuran (THF), Ethylenediaminetetraacetic acid (EDTA), hydrochloric acid, sulfuric acid, fibrinogen from bovine plasma, and N, N -dimethylacetamide (DMAc) were obtained from Sigma-Aldrich (St. Louis, MO).

    Purification:

    Article Title: Pharmacological characterization of the seven human NOX isoforms and their inhibitors
    Article Snippet: .. Horseradish peroxidase (HRP), flavin adenine dinucleotide disodium salt hydrate (FAD-Na2 ), phorbol 12-myristate 13-acetate (PMA), ionomycin calcium, superoxide dismutase (SOD), catalase, purified myeloperoxidase, dimethyl sulfoxide (DMSO), hydrogen peroxide 35% (H2 O2 ), 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), ethylenedinitrilotetraacetic acid (EDTA), N,N-bis(carboxymethyl)glycine (NTA), 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N'-tetraacetic acid (BAPTA), 3-sn -phosphatidic acid sodium salt, tetramethyl benzidine, dimethylformamide, MgCl2 , VAS2870 (CAS 722456-31-7), ebselen (CAS 60940-34-3), diapocynin (CAS 29799-22-2) and apocynin (CAS 498-02-2) were purchased from Sigma-Aldrich. ..

    Article Title: Highly Sensitive Genosensing Coupling Rolling Circle Amplification with Multiple DNAzyme Cores for DNA Walking.
    Article Snippet: Herein, a highly sensitive electrochemical genosensor is proposed by the construction of an innovative DNA walking machine. .. Herein, a highly sensitive electrochemical genosensor is proposed by the construction of an innovative DNA walking machine.

    Article Title: Development of an LC-MS/MS peptide mapping protocol for the NISTmAb
    Article Snippet: .. Guanidine HCl (Cat #RDD001), Tris(hydroxymethyl)aminomethane (Cat #T6066), Tris(hydroxymethyl)aminomethane HCl (Cat #T5941), ethylenediaminetetraacetic acid (EDTA) (Cat #39692), urea (Cat #U0631), acetic acid (Cat #695084), iodoacetamide (IAM) (Cat #A3221), recombinant porcine trypsin expressed in P. pastoris (Cat #03708985001), trypsin purified from bovine pancreas (Cat #TRYPSEQM-RO) and trypsin purified from porcine pancreas (Cat #T6567) were purchased from Sigma Aldrich. .. Recombinant bovine trypsin expressed in corn (Cat #PRO-313) and recombinant human-2 trypsin expressed in E. coli (Cat # PRO-770) were purchased from ProSpec.

    Impedance Spectroscopy:

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO). .. A CHI 660C Electrochemical Workstation (CH Instrument, Inc., Austin, TX) was used for DPV and EIS investigations.

    Article Title: Application of Bioconjugation Chemistry on Biosensor Fabrication for Detection of TAR-DNA binding Protein 43
    Article Snippet: Monoclonal Anti-TARDBP antibody produced in mouse (Cat. WH0023435M1), phosphate-buffer saline (PBS) 1.0 M (pH 7.4), undiluted human serum, ethylenediaminetetraacetic acid (EDTA) (Cat. EDS), 11-Mercaptoundecanoic acid (11-MUA) (Cat. 450561), 3-Mercaptopropionic acid (3-MPA) (Cat. M5801), N-(3 dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), and N–hydroxysuccinimide (NHS), dimethyl sulfoxide (DMSO), Amicon ultra-15 10K and amicon ultra-0.5 10K filters were purchased from Sigma Aldrich (St. Louis, MO). .. A CHI 660C Electrochemical Workstation (CH Instrument, Inc., Austin, TX) was used for DPV and EIS investigations.

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Development of an LC-MS/MS peptide mapping protocol for the NISTmAb
    Article Snippet: Guanidine HCl (Cat #RDD001), Tris(hydroxymethyl)aminomethane (Cat #T6066), Tris(hydroxymethyl)aminomethane HCl (Cat #T5941), ethylenediaminetetraacetic acid (EDTA) (Cat #39692), urea (Cat #U0631), acetic acid (Cat #695084), iodoacetamide (IAM) (Cat #A3221), recombinant porcine trypsin expressed in P. pastoris (Cat #03708985001), trypsin purified from bovine pancreas (Cat #TRYPSEQM-RO) and trypsin purified from porcine pancreas (Cat #T6567) were purchased from Sigma Aldrich. .. Dithiothreitol (DTT) (Cat #20291), Zeba™ Spin 7 K MWCO size-exclusion desalting columns (Cat #89882), LC/MS grade water (Cat # W6212), 0.1% formic acid in water (Cat # LS118) and 0.1% formic acid in acetonitrile (Cat #LS120) were purchased from Fisher Scientific.

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    Millipore pbs edta
    Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as <t>PBS-EDTA,</t> to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.
    Pbs Edta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs edta/product/Millipore
    Average 99 stars, based on 23 article reviews
    Price from $9.99 to $1999.99
    pbs edta - by Bioz Stars, 2020-03
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    99
    Millipore calpain reaction buffer
    Schematic diagram showing the relationship of <t>calpain/NF-κB/inflammation/NVU</t> damage after CCI in mice. Traumatic brain injury induces calcium overload, which, in turn, upregulates calpain. Calpain may downregulate IκB and activate NF-κB. NF-κB induces activation of TNF-α, iNOS, ICAM-1, and MMP-9. These inflammatory substances induce degradation of basal lamina and tight junction proteins, resulting in NVU disruption, leading to brain edema. MDL28170 could reverse those changes. NF-κB: Nuclear factor-κB; NVU: Neurovascular unit; CCI: Controlled cortical impact; IκB: Inhibitory-κB; TNF-α: Tumor necrosis factor-α; iNOS: Inducible nitric oxide synthase; ICAM-1: Intracellular adhesion molecule-1; MMP-9: Matrix metalloproteinase-9.
    Calpain Reaction Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/calpain reaction buffer/product/Millipore
    Average 99 stars, based on 6 article reviews
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    99
    Millipore m edta
    Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with <t>EDTA,</t> <t>EGTA</t>
    M Edta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m edta/product/Millipore
    Average 99 stars, based on 2 article reviews
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    99
    Millipore immunoprecipitation buffer
    Binding of Wt1 protein to the 5′-flanking region of the Adamts16 gene. ChIP was performed to detect Wt1 protein bound to the 5′-flanking region of the Adamts16 gene in its native chromosomal configuration. The drawing ( A ) delineates the three predicted Wt1 binding sites ( Wt1-A , Wt1-B , and Wt1-C ) in the promoter and 5′-UTR of the Adamts16 gene and allocates the PCR primers used for DNA amplification. Specific antibodies against Wt1 and histone proteins were chosen for <t>immunoprecipitation</t> of M15 whole cell lysates. Amplicons encompassing the 5′-flanking region of the Adamts16 gene were enriched ∼2.5-fold with the use of anti-Wt1 antibody compared with normal rabbit IgG (NRb-IgG). No differences in actin DNA were observed between anti-Wt1 antibody and normal rabbit IgG ( B ). The gene encoding anti-Müllerian hormone receptor 2 ( Amhr2 ), served as a positive control ( B ). Binding of Wt1(−KTS) protein to the Adamts16 promoter was confirmed in stimulated UB27 cells ( C ). Wt1(+KTS) protein failed to interact with the promoter of the Adamts16 gene in UD28 cells ( D ). Data shown are means ± S.E. ( error bars ). Statistical significances are indicated by asterisks (*, p
    Immunoprecipitation Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as PBS-EDTA, to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as PBS-EDTA, to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Concentration Assay, Buffer Exchange

    (A) Reverse phase chromatogram of AMB-PEG and unconjugated AMB, with eluted peaks detected at 406 nm. 50 μL of AMB-PEG 1 and 2, and 10 μL of unconjugated AMB dispersed in PBS-EDTA and 48% ACN respectively were injected into a C18 reverse phase column and eluted isocratically in a 48% ACN buffer at a flow rate of 0.5 ml/min for 40 minutes. Peaks were detected at 406 nm. The AMB-PEG conjugate has a shorter retention time, implying that it is more hydrophilic. From the AMB-PEG samples, AMB-PEG conjugate and free AMB (based on the retention time of unconjugated AMB) fractions were collected for further analysis via size exclusion chromatography. (B) Size exclusion chromatogram of AMB-PEG 2 and unconjugated AMB, as well as the relevant fractions collected from RPC, in a 20% ACN mobile phase. 20 mM of AMB-PEG 2 formulations and unconjugated AMB in DMSO were prepared and resuspended at 2 mM in a 20% ACN buffer. 50 μL of these samples (unconjugated AMB diluted tenfold prior to analysis) and the peak fractions collected previously from RPC were passed through a size exclusion column and eluted peaks were detected at 406 nm. Unconjugated AMB was eluted later compared to the AMB-PEG conjugate, implying that it has a smaller hydrodynamic volume compared to AMB-PEG in 20% ACN. The previously collected AMB-PEG conjugate and free AMB peak fractions had similar retention times as the AMB-PEG 2 formulation and unconjugated AMB samples respectively, thus verifying their respective peak identities. AMB-PEG 1 and 2 have identical elution profiles. (C) Size exclusion chromatogram of AMB-PEG and unconjugated AMB dispersed in PBS-EDTA. 3 μL of 2 mM AMB-PEG formulations that have been retained by 10 kDa centrifugal filters (Millipore) and 50 μL of the supernatant obtained from unconjugated AMB were analysed using a Superdex 75 size exclusion column and eluted peaks detected at 406 nm. In a PBS-EDTA mobile phase, unconjugated AMB has a shorter retention time of 20 minutes compared to AMB-PEG at 40 minutes, implying that AMB-PEG has a smaller hydrodynamic volume under these experimental conditions.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: (A) Reverse phase chromatogram of AMB-PEG and unconjugated AMB, with eluted peaks detected at 406 nm. 50 μL of AMB-PEG 1 and 2, and 10 μL of unconjugated AMB dispersed in PBS-EDTA and 48% ACN respectively were injected into a C18 reverse phase column and eluted isocratically in a 48% ACN buffer at a flow rate of 0.5 ml/min for 40 minutes. Peaks were detected at 406 nm. The AMB-PEG conjugate has a shorter retention time, implying that it is more hydrophilic. From the AMB-PEG samples, AMB-PEG conjugate and free AMB (based on the retention time of unconjugated AMB) fractions were collected for further analysis via size exclusion chromatography. (B) Size exclusion chromatogram of AMB-PEG 2 and unconjugated AMB, as well as the relevant fractions collected from RPC, in a 20% ACN mobile phase. 20 mM of AMB-PEG 2 formulations and unconjugated AMB in DMSO were prepared and resuspended at 2 mM in a 20% ACN buffer. 50 μL of these samples (unconjugated AMB diluted tenfold prior to analysis) and the peak fractions collected previously from RPC were passed through a size exclusion column and eluted peaks were detected at 406 nm. Unconjugated AMB was eluted later compared to the AMB-PEG conjugate, implying that it has a smaller hydrodynamic volume compared to AMB-PEG in 20% ACN. The previously collected AMB-PEG conjugate and free AMB peak fractions had similar retention times as the AMB-PEG 2 formulation and unconjugated AMB samples respectively, thus verifying their respective peak identities. AMB-PEG 1 and 2 have identical elution profiles. (C) Size exclusion chromatogram of AMB-PEG and unconjugated AMB dispersed in PBS-EDTA. 3 μL of 2 mM AMB-PEG formulations that have been retained by 10 kDa centrifugal filters (Millipore) and 50 μL of the supernatant obtained from unconjugated AMB were analysed using a Superdex 75 size exclusion column and eluted peaks detected at 406 nm. In a PBS-EDTA mobile phase, unconjugated AMB has a shorter retention time of 20 minutes compared to AMB-PEG at 40 minutes, implying that AMB-PEG has a smaller hydrodynamic volume under these experimental conditions.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Injection, Flow Cytometry, Size-exclusion Chromatography

    (A) AMB-PEG reaction scheme. The NHS ester on MS(PEG) 4 reacts with the primary amine (–NH2) group on AMB at a 1:1 molar ratio, generating conjugated AMB-PEG via amide bond formation. (B) Solubility of AMB-PEG at varying AMB:PEG molar ratios. AMB-PEG mixtures and unreacted AMB were prepared in DMSO, incubated for 2 hours and then (i) dispersed to a final concentration of 2 mM in PBS-EDTA, containing 10% DMSO by volume. (ii) The mixtures were then centrifuged to facilitate the observation of insoluble precipitates. Higher molar ratios yielded a suspension of yellow particles that precipitated upon centrifugation, similar to what was observed with unconjugated AMB.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: (A) AMB-PEG reaction scheme. The NHS ester on MS(PEG) 4 reacts with the primary amine (–NH2) group on AMB at a 1:1 molar ratio, generating conjugated AMB-PEG via amide bond formation. (B) Solubility of AMB-PEG at varying AMB:PEG molar ratios. AMB-PEG mixtures and unreacted AMB were prepared in DMSO, incubated for 2 hours and then (i) dispersed to a final concentration of 2 mM in PBS-EDTA, containing 10% DMSO by volume. (ii) The mixtures were then centrifuged to facilitate the observation of insoluble precipitates. Higher molar ratios yielded a suspension of yellow particles that precipitated upon centrifugation, similar to what was observed with unconjugated AMB.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Mass Spectrometry, Solubility, Incubation, Concentration Assay, Centrifugation

    Aqueous solubility of AMB-PEG 1 and 2. Increasing amounts of AMB-PEG formulations were added to PBS-EDTA, and post-centrifugation, the concentration of soluble AMB-PEG in the supernatants quantified based on their absorbance at 365 nm. Error bars denote the standard deviation between 3 independent formulations. Dotted line represents the theoretical condition where AMB formulation is completely soluble (i.e. concentration of total AMB = concentration of soluble AMB).

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: Aqueous solubility of AMB-PEG 1 and 2. Increasing amounts of AMB-PEG formulations were added to PBS-EDTA, and post-centrifugation, the concentration of soluble AMB-PEG in the supernatants quantified based on their absorbance at 365 nm. Error bars denote the standard deviation between 3 independent formulations. Dotted line represents the theoretical condition where AMB formulation is completely soluble (i.e. concentration of total AMB = concentration of soluble AMB).

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Solubility, Centrifugation, Concentration Assay, Standard Deviation

    Schematic diagram showing the relationship of calpain/NF-κB/inflammation/NVU damage after CCI in mice. Traumatic brain injury induces calcium overload, which, in turn, upregulates calpain. Calpain may downregulate IκB and activate NF-κB. NF-κB induces activation of TNF-α, iNOS, ICAM-1, and MMP-9. These inflammatory substances induce degradation of basal lamina and tight junction proteins, resulting in NVU disruption, leading to brain edema. MDL28170 could reverse those changes. NF-κB: Nuclear factor-κB; NVU: Neurovascular unit; CCI: Controlled cortical impact; IκB: Inhibitory-κB; TNF-α: Tumor necrosis factor-α; iNOS: Inducible nitric oxide synthase; ICAM-1: Intracellular adhesion molecule-1; MMP-9: Matrix metalloproteinase-9.

    Journal: Chinese Medical Journal

    Article Title: Protective Effects of Calpain Inhibition on Neurovascular Unit Injury through Downregulating Nuclear Factor-κB-related Inflammation during Traumatic Brain Injury in Mice

    doi: 10.4103/0366-6999.198001

    Figure Lengend Snippet: Schematic diagram showing the relationship of calpain/NF-κB/inflammation/NVU damage after CCI in mice. Traumatic brain injury induces calcium overload, which, in turn, upregulates calpain. Calpain may downregulate IκB and activate NF-κB. NF-κB induces activation of TNF-α, iNOS, ICAM-1, and MMP-9. These inflammatory substances induce degradation of basal lamina and tight junction proteins, resulting in NVU disruption, leading to brain edema. MDL28170 could reverse those changes. NF-κB: Nuclear factor-κB; NVU: Neurovascular unit; CCI: Controlled cortical impact; IκB: Inhibitory-κB; TNF-α: Tumor necrosis factor-α; iNOS: Inducible nitric oxide synthase; ICAM-1: Intracellular adhesion molecule-1; MMP-9: Matrix metalloproteinase-9.

    Article Snippet: [ ] In brief, cytosolic and mitochondrial proteins (30 μg) were incubated with calpain reaction buffer (20 mmol/L HEPES, 1 mmol/L EDTA, 50 mmol/L NaCl, and 0.1% (v/v) 2-mercaptoethanol, containing 10 μmol/L calpain I fluorescent substrate [Calbiochem Co., La Jolla, CA, USA], pH 7.6).

    Techniques: Mouse Assay, Activation Assay

    MDL28170 treatment suppresses the calpain activity in the cytosolic and mitochondrial fractions and upregulates the expression of calpastatin in the cytosolic fractions. (a and b) The bar graphs reflect the calpain activity in the cytosolic fractions and mitochondrial fractions at 6 h and 24 h. (c) Representative Western blots of calpastatin and β-actin from each group; (d) the results were quantified and are shown as the mean ± SD. * P

    Journal: Chinese Medical Journal

    Article Title: Protective Effects of Calpain Inhibition on Neurovascular Unit Injury through Downregulating Nuclear Factor-κB-related Inflammation during Traumatic Brain Injury in Mice

    doi: 10.4103/0366-6999.198001

    Figure Lengend Snippet: MDL28170 treatment suppresses the calpain activity in the cytosolic and mitochondrial fractions and upregulates the expression of calpastatin in the cytosolic fractions. (a and b) The bar graphs reflect the calpain activity in the cytosolic fractions and mitochondrial fractions at 6 h and 24 h. (c) Representative Western blots of calpastatin and β-actin from each group; (d) the results were quantified and are shown as the mean ± SD. * P

    Article Snippet: [ ] In brief, cytosolic and mitochondrial proteins (30 μg) were incubated with calpain reaction buffer (20 mmol/L HEPES, 1 mmol/L EDTA, 50 mmol/L NaCl, and 0.1% (v/v) 2-mercaptoethanol, containing 10 μmol/L calpain I fluorescent substrate [Calbiochem Co., La Jolla, CA, USA], pH 7.6).

    Techniques: Activity Assay, Expressing, Western Blot

    Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with EDTA, EGTA

    Journal: Immunology

    Article Title: Natural lysophospholipids reduce Mycobacterium tuberculosis-induced cytotoxicity and induce anti-mycobacterial activity by a phagolysosome maturation-dependent mechanism in A549 type II alveolar epithelial cells

    doi: 10.1111/j.1365-2567.2009.03145.x

    Figure Lengend Snippet: Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with EDTA, EGTA

    Article Snippet: In several experiments, 2 m m EDTA or 3 m m EGTA (Calbiochem) were added 15 min before the addition of LPA or S1P, whereas 20 μ m 1,2-bis(2-aminophenoxy)ethane-N,N,N1,N1-tetraacetic acid tetraicis (acetoxymethyl ester) (BAPTA-AM) (Sigma-Aldrich, Milan, Italy) was added 30 min before stimulation with LPA or S1P.

    Techniques: Activation Assay

    Binding of Wt1 protein to the 5′-flanking region of the Adamts16 gene. ChIP was performed to detect Wt1 protein bound to the 5′-flanking region of the Adamts16 gene in its native chromosomal configuration. The drawing ( A ) delineates the three predicted Wt1 binding sites ( Wt1-A , Wt1-B , and Wt1-C ) in the promoter and 5′-UTR of the Adamts16 gene and allocates the PCR primers used for DNA amplification. Specific antibodies against Wt1 and histone proteins were chosen for immunoprecipitation of M15 whole cell lysates. Amplicons encompassing the 5′-flanking region of the Adamts16 gene were enriched ∼2.5-fold with the use of anti-Wt1 antibody compared with normal rabbit IgG (NRb-IgG). No differences in actin DNA were observed between anti-Wt1 antibody and normal rabbit IgG ( B ). The gene encoding anti-Müllerian hormone receptor 2 ( Amhr2 ), served as a positive control ( B ). Binding of Wt1(−KTS) protein to the Adamts16 promoter was confirmed in stimulated UB27 cells ( C ). Wt1(+KTS) protein failed to interact with the promoter of the Adamts16 gene in UD28 cells ( D ). Data shown are means ± S.E. ( error bars ). Statistical significances are indicated by asterisks (*, p

    Journal: The Journal of Biological Chemistry

    Article Title: Transcriptional Regulation by the Wilms Tumor Protein, Wt1, Suggests a Role of the Metalloproteinase Adamts16 in Murine Genitourinary Development *

    doi: 10.1074/jbc.M113.464644

    Figure Lengend Snippet: Binding of Wt1 protein to the 5′-flanking region of the Adamts16 gene. ChIP was performed to detect Wt1 protein bound to the 5′-flanking region of the Adamts16 gene in its native chromosomal configuration. The drawing ( A ) delineates the three predicted Wt1 binding sites ( Wt1-A , Wt1-B , and Wt1-C ) in the promoter and 5′-UTR of the Adamts16 gene and allocates the PCR primers used for DNA amplification. Specific antibodies against Wt1 and histone proteins were chosen for immunoprecipitation of M15 whole cell lysates. Amplicons encompassing the 5′-flanking region of the Adamts16 gene were enriched ∼2.5-fold with the use of anti-Wt1 antibody compared with normal rabbit IgG (NRb-IgG). No differences in actin DNA were observed between anti-Wt1 antibody and normal rabbit IgG ( B ). The gene encoding anti-Müllerian hormone receptor 2 ( Amhr2 ), served as a positive control ( B ). Binding of Wt1(−KTS) protein to the Adamts16 promoter was confirmed in stimulated UB27 cells ( C ). Wt1(+KTS) protein failed to interact with the promoter of the Adamts16 gene in UD28 cells ( D ). Data shown are means ± S.E. ( error bars ). Statistical significances are indicated by asterisks (*, p

    Article Snippet: The supernatants were diluted in immunoprecipitation buffer (0.01% SDS, 1.1% Triton X-100, 1.2 m m EDTA, 16.7 m m Tris-HCl, pH 8.1) and precleared for 1 h at 4 °C with DNA-blocked protein G-agarose (Millipore).

    Techniques: Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Amplification, Immunoprecipitation, Positive Control