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Applichem ethylenediaminetetraacetic acid
Ethylenediaminetetraacetic Acid, supplied by Applichem, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ethylenediaminetetraacetic acid - by Bioz Stars, 2020-03
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Positive Control:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: The RNA probe synthesized on the Krox20 gene template, whose expression level is high in the central nervous system cells of mouse embryos at these stages of development, was used as a positive control. .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich).

Synthesized:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: The RNA probe synthesized on the Krox20 gene template, whose expression level is high in the central nervous system cells of mouse embryos at these stages of development, was used as a positive control. .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich).

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: DIG-labeled antisense RNA probes for in situ hybridization were synthesized by in vitro transcription. .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Cytometry:

Article Title: Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis
Article Snippet: Paragraph title: Flow cytometry of S1P1 + T and B cells in lymph nodes and blood ... Blood was obtained by cardiac puncture, and 5 μl of 0.5 M EDTA (AppliChem) was added.

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: Samples and single color controls were incubated at 4°C, washed and analyzed by flow cytometry using a BD LSRII flow cytometer. .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

Blocking Assay:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich). .. The embryos were incubated at room temperature in a (1 : 1) mixture of a hybridization buffer and PBS and then in the hybridization buffer.

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: For the hematopoietic staining, cells were first incubated with 10% human serum (Lonza, Basel, Switzerland) to block unspecific binding, then the following antibodies were added: antihematopoietic lineage eFluor 450 cocktail (eBioscience, San Diego, Calif), CD34 APC (BD Pharmingen, Franklin Lakes, NJ), CD38 PE (BD Pharmingen), CD90 Clone 5E10 Pe-Cy7 (BD Pharmingen), CD123 Clone 7G3 PerCP-Cy5.5 (BD Pharmingen), and CD45RA FITC Ref. A07786 (BeckmanCoulter, Brea, Calif) (all antihuman). .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Cells were collected in FACS buffer (PBS/0.1% FCS/1 mM EDTA) and resuspended at a cell concentration of 5 × 105 cells/100 μl and incubated with 2 μg/ml of human IgG for 45 min at 4°C to block unspecific binding of antibodies to Fcγ receptors.

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS. .. Subsequently, sections were washed in PBS, post-fixed in 4% PFA containing 0.2% glutaraldehyde (Sigma) in PBS for 15 min, washed in PBS, and prehybridized for 2 h at 68°C in hybridization buffer containing 50% deionized formamide (AppliChem), 5× SSC, 1% blocking reagent (Roche), 5 mM EDTA, 0.1% Tween 20 (Sigma), 0.1% CHAPS (Sigma), 0.1 mg/ml heparin (Sigma), and 100 µg/ml yeast RNA (Invitrogen).Sections were then hybridized overnight at 68°C in the same buffer containing denatured probes.

Incubation:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich). .. The embryos were incubated at room temperature in a (1 : 1) mixture of a hybridization buffer and PBS and then in the hybridization buffer.

Article Title: Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis
Article Snippet: Blood was obtained by cardiac puncture, and 5 μl of 0.5 M EDTA (AppliChem) was added. .. All samples were incubated with BD Horizon™ FVS450 (BD Biosciences) at 4 °C for 30 min.

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: Samples and single color controls were incubated at 4°C, washed and analyzed by flow cytometry using a BD LSRII flow cytometer. .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: .. After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Cells were collected in FACS buffer (PBS/0.1% FCS/1 mM EDTA) and resuspended at a cell concentration of 5 × 105 cells/100 μl and incubated with 2 μg/ml of human IgG for 45 min at 4°C to block unspecific binding of antibodies to Fcγ receptors.

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS. .. Subsequently, sections were washed in PBS, post-fixed in 4% PFA containing 0.2% glutaraldehyde (Sigma) in PBS for 15 min, washed in PBS, and prehybridized for 2 h at 68°C in hybridization buffer containing 50% deionized formamide (AppliChem), 5× SSC, 1% blocking reagent (Roche), 5 mM EDTA, 0.1% Tween 20 (Sigma), 0.1% CHAPS (Sigma), 0.1 mg/ml heparin (Sigma), and 100 µg/ml yeast RNA (Invitrogen).Sections were then hybridized overnight at 68°C in the same buffer containing denatured probes.

Expressing:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: The RNA probe synthesized on the Krox20 gene template, whose expression level is high in the central nervous system cells of mouse embryos at these stages of development, was used as a positive control. .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: Paragraph title: Surface Receptor Expression ... After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C.

Article Title: Ginkgolic Acid is a Multi-Target Inhibitor of Key Enzymes in Pro-Inflammatory Lipid Mediator Biosynthesis
Article Snippet: .. Zileuton [N -(1-benzo[b]thien-2-ylethyl)-N -hydroxyurea] was from Sequoia Research Products (Oxford, UK); PGH2 was from Larodan (Malmö, Sweden); IL-1β was from ReproTech (Hamburg, Germany); RSC-3388 was from Calbiochem (Darmstadt, Germany); EDTA and Nonidet P-40 were from AppliChem (Darmstadt, Germany); p-anisidinium chloride was from Merck (Darmstadt, Germany); Insect Express Sf9-S2 and RPMI media, glutamine, penicillin, and streptomycin were from PAA (Coelbe, Germany); Bac-to-Bac baculovirus expression system was from Invitrogen (Karlsruhe, Germany); Ni-NTA agarose was from Qiagen (Hilden, Germany). .. AA, Ca2+ -ionophore A23187, dextrane, dithiothreitol, fetal calf serum, indomethacin, lipopolysaccharide (LPS), Triton X-100, and all other chemicals were purchased from Sigma-Aldrich (Taufkirchen, Germany), unless stated otherwise.

Modification:

Article Title: Decisive role of P42/44 mitogen-activated protein kinase in Δ9-tetrahydrocannabinol-induced migration of human mesenchymal stem cells
Article Snippet: Dulbecco's Modified Eagle's medium (DMEM, high glucose, GlutaMAXTM ), trypsin-EDTA and penicillin-streptomycin were obtained from Life Technologies GmbH (Darmstadt, Germany). .. Dimethyl sulfoxide (DMSO), EDTA, glycerol, glycine, H2 O2 , HEPES, MgCl2 , NaCl, sodium dodecyl sulphate and Tris were bought from AppliChem GmbH (Darmstadt, Germany).

Western Blot:

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis
Article Snippet: .. Western Blot Proteins were isolated from wild-type and TzfpGTi/GTi testis using RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 0.1% SDS, 0.5 mM PMSF (AppliChem) and 1× Protease Inhibitor Coctail (Sigma)). .. The protein concentration was measured using Protein assay dye reagent concentrate (BioRad) and 150 µg protein was loaded onto NuPAGE® 10% Bis-Tris Gel 1.0 mm (Invitrogen).

Derivative Assay:

Article Title: Loss of menin in osteoblast lineage affects osteocyte–osteoclast crosstalk causing osteoporosis
Article Snippet: Osteocytes were extracted by sequential digestions with collagenase and EDTA (AppliChem, Darmstadt, Germany). .. Fractions 7–9 and cells derived from bone particles were pooled and considered as osteocytes.

Hybridization:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich). .. The embryos were incubated at room temperature in a (1 : 1) mixture of a hybridization buffer and PBS and then in the hybridization buffer.

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS. .. Subsequently, sections were washed in PBS, post-fixed in 4% PFA containing 0.2% glutaraldehyde (Sigma) in PBS for 15 min, washed in PBS, and prehybridized for 2 h at 68°C in hybridization buffer containing 50% deionized formamide (AppliChem), 5× SSC, 1% blocking reagent (Roche), 5 mM EDTA, 0.1% Tween 20 (Sigma), 0.1% CHAPS (Sigma), 0.1 mg/ml heparin (Sigma), and 100 µg/ml yeast RNA (Invitrogen).Sections were then hybridized overnight at 68°C in the same buffer containing denatured probes.

High Performance Liquid Chromatography:

Article Title: Ginkgolic Acid is a Multi-Target Inhibitor of Key Enzymes in Pro-Inflammatory Lipid Mediator Biosynthesis
Article Snippet: Zileuton [N -(1-benzo[b]thien-2-ylethyl)-N -hydroxyurea] was from Sequoia Research Products (Oxford, UK); PGH2 was from Larodan (Malmö, Sweden); IL-1β was from ReproTech (Hamburg, Germany); RSC-3388 was from Calbiochem (Darmstadt, Germany); EDTA and Nonidet P-40 were from AppliChem (Darmstadt, Germany); p-anisidinium chloride was from Merck (Darmstadt, Germany); Insect Express Sf9-S2 and RPMI media, glutamine, penicillin, and streptomycin were from PAA (Coelbe, Germany); Bac-to-Bac baculovirus expression system was from Invitrogen (Karlsruhe, Germany); Ni-NTA agarose was from Qiagen (Hilden, Germany). .. HPLC/UPLC solvents were from VWR (Darmstadt, Germany).

Flow Cytometry:

Article Title: Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis
Article Snippet: Paragraph title: Flow cytometry of S1P1 + T and B cells in lymph nodes and blood ... Blood was obtained by cardiac puncture, and 5 μl of 0.5 M EDTA (AppliChem) was added.

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: Samples and single color controls were incubated at 4°C, washed and analyzed by flow cytometry using a BD LSRII flow cytometer. .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

Protease Inhibitor:

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis
Article Snippet: .. Western Blot Proteins were isolated from wild-type and TzfpGTi/GTi testis using RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 0.1% SDS, 0.5 mM PMSF (AppliChem) and 1× Protease Inhibitor Coctail (Sigma)). .. The protein concentration was measured using Protein assay dye reagent concentrate (BioRad) and 150 µg protein was loaded onto NuPAGE® 10% Bis-Tris Gel 1.0 mm (Invitrogen).

Cell Culture:

Article Title: Oxidative status predicts quality in human mesenchymal stem cells
Article Snippet: MSC-collagen constructs were cultured for 3 weeks in chondrogenic medium consisting of advanced DMEM + GlutaMAX (Gibco), 2.5% FBS, 100 units/mL penicillin, 100 mg/mL streptomycin, 2.5 μg/mL amphotericin B, 40 ng/mL dexamethasone (Sigma), 50 μg/mL ascorbic acid 2-phosphate (Sigma), 35 μg/mL l -proline (Sigma), 1× Insulin-Transferrin-Selenium X (Gibco), and 10 ng/ml transforming growth factor (TGF)-β1 (Peprotech). .. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l -cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem).

Cell Differentiation:

Article Title: Oxidative status predicts quality in human mesenchymal stem cells
Article Snippet: Chondrogenic differentiation Collagen type I cubes (Biopad, Euroresearch, Italy) were used as a scaffold to support cellular differentiation [ ]. .. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l -cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem).

other:

Article Title: Spatial considerations during cryopreservation of a large volume sample
Article Snippet: 2.9.2 Total cell counts A known volume of ELS was removed from alginate post-cryopreservation in 16 mM EDTA (Applichem, Darmstadt, Germany) solution before the ELS were disaggregated and a nucleic count carried out using the nucleocounter system.

Article Title: PMMA Third-Body Wear after Unicondylar Knee Arthroplasty Decuples the UHMWPE Wear Particle Generation In Vitro
Article Snippet: The applied lubricant was changed every six days (25% (v/v) newborn calf serum (Biochrom, Germany) with 0.1% (m/v) sodium azide solution in sterile water with EDTA (AppliChem, Germany) for pH stability and Amphotericin B (Biochrom, Germany) as a fungicidal).

Article Title: Inhibition of FAAH confers increased stem cell migration via PPARα
Article Snippet: Bisbenzimide, DMSO, EDTA, glycerol, glycine, H2 O2 , HEPES, NaCl, SDS, and Tris were obtained from AppliChem GmbH (Darmstadt, Germany).

Protein Concentration:

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis
Article Snippet: Western Blot Proteins were isolated from wild-type and TzfpGTi/GTi testis using RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 0.1% SDS, 0.5 mM PMSF (AppliChem) and 1× Protease Inhibitor Coctail (Sigma)). .. The protein concentration was measured using Protein assay dye reagent concentrate (BioRad) and 150 µg protein was loaded onto NuPAGE® 10% Bis-Tris Gel 1.0 mm (Invitrogen).

Binding Assay:

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: For the hematopoietic staining, cells were first incubated with 10% human serum (Lonza, Basel, Switzerland) to block unspecific binding, then the following antibodies were added: antihematopoietic lineage eFluor 450 cocktail (eBioscience, San Diego, Calif), CD34 APC (BD Pharmingen, Franklin Lakes, NJ), CD38 PE (BD Pharmingen), CD90 Clone 5E10 Pe-Cy7 (BD Pharmingen), CD123 Clone 7G3 PerCP-Cy5.5 (BD Pharmingen), and CD45RA FITC Ref. A07786 (BeckmanCoulter, Brea, Calif) (all antihuman). .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Cells were collected in FACS buffer (PBS/0.1% FCS/1 mM EDTA) and resuspended at a cell concentration of 5 × 105 cells/100 μl and incubated with 2 μg/ml of human IgG for 45 min at 4°C to block unspecific binding of antibodies to Fcγ receptors.

Article Title: Oxidative status predicts quality in human mesenchymal stem cells
Article Snippet: .. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l -cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem). .. GAG accumulation was determined by binding to alcian blue (Fluka, Sigma) and quantified using chondroitin sulphate (Sigma) reference standards [ ].

Staining:

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany). .. Antibodies were added as follows: CD14/CD20/CD34/CD45 Vioblue (all Miltenyi Biotec, Bergisch Gladbach, Germany), CD73 APC (Miltenyi Biotec), CD105 PE (Mitenyi Biotec), and CD90 Clone 5E10 Pe-Cy7 (BD Pharmingen) (all antihuman).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Staining was performed for 30 min at 4°C in the dark in PBS using the following antibodies: anti-MHC II-FITC (Immunotools), anti-tyrosine-protein-kinase Mer (MerTK)-PE (R & D Systems, Minneapolis, MN, USA), anti-programmed death ligand 1 (PD-L1/CD274)-APC and anti-CD14-PeCy7 (both from Biolegend, San Diego, CA, USA) (antibody panel 1), or anti-CD86-FITC (Biolegend), anti-lipoprotein receptor-related protein 1 (LRP-1/CD91)-PE (Thermo Fisher Scientific, Waltham, MA, USA), anti-leukocyte-associated immunoglobulin-like receptor 1 (LAIR1/CD305)-AF647 and anti-scavenger receptor A 1 (SR-A1/CD204)-PeCy7 (both from Biolegend) (antibody panel 2).

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Paragraph title: Tissue preparation, immunohistochemistry, X-Gal staining, and in situ hybridization ... Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Nucleic Acid Electrophoresis:

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Plasmids carrying subcloned coding regions of AS021 surrounding genes were linearized by restriction digestion and incubated for 3 h at 37°C in a 20-µl reaction mixture containing 2 µg linearized template plasmids, 2 µl 10× transcription buffer (Roche), 2 µl DIG-labeled RNA mix (Roche), 0.5 µl RNase inhibitor (New England Biolabs), 2 µl T7 or Sp6 RNA polymerase (Roche), and DEPC-treated H2 O. cRNA probes were purified by lithium chloride precipitation and monitored by gel electrophoresis. .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Isolation:

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis
Article Snippet: .. Western Blot Proteins were isolated from wild-type and TzfpGTi/GTi testis using RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 0.1% SDS, 0.5 mM PMSF (AppliChem) and 1× Protease Inhibitor Coctail (Sigma)). .. The protein concentration was measured using Protein assay dye reagent concentrate (BioRad) and 150 µg protein was loaded onto NuPAGE® 10% Bis-Tris Gel 1.0 mm (Invitrogen).

Article Title: Loss of menin in osteoblast lineage affects osteocyte–osteoclast crosstalk causing osteoporosis
Article Snippet: Paragraph title: Osteocyte isolation ... Osteocytes were extracted by sequential digestions with collagenase and EDTA (AppliChem, Darmstadt, Germany).

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Mouse brains were isolated, fixed for 3–4 h in 5% formalin in aPB containing 25% sucrose, rinsed in H2 O, and embedded in Tissue-Tek O.C.T.™ Compound (Sakura). .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Immunohistochemistry:

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Paragraph title: Tissue preparation, immunohistochemistry, X-Gal staining, and in situ hybridization ... Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Mouse Assay:

Article Title: Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis
Article Snippet: Flow cytometry of S1P1 + T and B cells in lymph nodes and blood Naïve female B6 mice were treated with 1 mg/kg BW FTY720 or vehicle solution for 10 consecutive days. .. Blood was obtained by cardiac puncture, and 5 μl of 0.5 M EDTA (AppliChem) was added.

Article Title: Loss of menin in osteoblast lineage affects osteocyte–osteoclast crosstalk causing osteoporosis
Article Snippet: Osteocyte isolation Primary osteocyte-enriched fractions from Men1 gtRosaCreERT2 mice were isolated as previously described. .. Osteocytes were extracted by sequential digestions with collagenase and EDTA (AppliChem, Darmstadt, Germany).

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: For in situ hybridization at E16 or P0, NMRI wild-type mice were retrocardially perfused using alkaline phosphate buffer (aPB; 0.1 M Na2 HPO4 , pH 9, Merck) containing 10% sucrose (Merck) followed by 5% formalin (diluted from 37% formalin, Merck) in aPB containing 25% sucrose. .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Construct:

Article Title: Oxidative status predicts quality in human mesenchymal stem cells
Article Snippet: .. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l -cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem). .. GAG accumulation was determined by binding to alcian blue (Fluka, Sigma) and quantified using chondroitin sulphate (Sigma) reference standards [ ].

Lysis:

Article Title: Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis
Article Snippet: Blood was obtained by cardiac puncture, and 5 μl of 0.5 M EDTA (AppliChem) was added. .. Erythrocyte lysis was performed using an ammonium chloride-based red blood cell lysis buffer.

Purification:

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Plasmids carrying subcloned coding regions of AS021 surrounding genes were linearized by restriction digestion and incubated for 3 h at 37°C in a 20-µl reaction mixture containing 2 µg linearized template plasmids, 2 µl 10× transcription buffer (Roche), 2 µl DIG-labeled RNA mix (Roche), 0.5 µl RNase inhibitor (New England Biolabs), 2 µl T7 or Sp6 RNA polymerase (Roche), and DEPC-treated H2 O. cRNA probes were purified by lithium chloride precipitation and monitored by gel electrophoresis. .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

SDS Page:

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis
Article Snippet: Western Blot Proteins were isolated from wild-type and TzfpGTi/GTi testis using RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 0.5% NP-40, 0.1% SDS, 0.5 mM PMSF (AppliChem) and 1× Protease Inhibitor Coctail (Sigma)). .. After SDS-PAGE, the proteins were blotted onto a PVDF membrane and the membrane was blocked in PBS with 5% skim milk before incubating with primary antibody over night at 4°C.

Recombinant:

Article Title: Amino acid conjugates of lithocholic acid as antagonists of the EphA2 receptor
Article Snippet: Leupeptin, aprotinin, NP40, tween20, BSA and salts for solutions were from Applichem; EDTA and sodium orthovanadate were from Sigma. .. Leupeptin, aprotinin, NP40, tween20, BSA and salts for solutions were from Applichem; EDTA and sodium orthovanadate were from Sigma.

In Vitro:

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: DIG-labeled antisense RNA probes for in situ hybridization were synthesized by in vitro transcription. .. Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

In Situ Hybridization:

Article Title: A Mammalian Conserved Element Derived from SINE Displays Enhancer Properties Recapitulating Satb2 Expression in Early-Born Callosal Projection Neurons
Article Snippet: Paragraph title: Tissue preparation, immunohistochemistry, X-Gal staining, and in situ hybridization ... Tissue sections were immediately dried under vacuum for 30 min, fixed in 4% PFA (Merck) in PBS pH 7.4 for 15 min, washed in PBS, and incubated with 20 µg/ml proteinase K (Merck) in 20 mM Tris, pH 7.5/1 mM EDTA, pH 8 for 2.5 min. Proteinase K was inactivated in 0.2% glycine (AppliChem) in PBS.

Concentration Assay:

Article Title: Detection of T-Cadherin Expression in Mouse Embryos
Article Snippet: Immediately prior to hybridization, the embryos were rehydrated in solutions of decreasing concentrations of methanol (75, 50, and 25%), washed three times in PBS, and treated with proteinase K at room temperature (Qiagen, at a concentration of 10 μg/mL in PBS). .. Next, the embryos were gradually transferred into a hybridization buffer containing 50% formamide (Sigma-Aldrich), 5× SSC buffer (stock solution: 20× SSC buffer containing 3 M NaCl, 0.3 M sodium citrate, pH 7.0), 0.1% Triton X-100 (DiaEm), 50 μg/mL heparin (Sigma-Aldrich), 1 mg/mL RNA from type IV yeast (Sigma-Aldrich), 5 mM EDTA (Applichem), 2% blocking solution (Roche), and 0.1% CHAPS (3-[( -cholamidopropyl)-dimethylammonio]- 1-propanesulfonate) (Sigma-Aldrich).

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Cells were collected in FACS buffer (PBS/0.1% FCS/1 mM EDTA) and resuspended at a cell concentration of 5 × 105 cells/100 μl and incubated with 2 μg/ml of human IgG for 45 min at 4°C to block unspecific binding of antibodies to Fcγ receptors.

Cell Counting:

Article Title: Distinct Dynamics of Stem and Progenitor Cells in Blood of Polytraumatized Patients
Article Snippet: Afterward cell count was taken and 2 × 106 cells were used for hematopoietic staining and mesenchymal staining respectively. .. For MSC staining cells were resuspended in PBS with 0.5% FCS and 2 mM EDTA (AppliChem, Darmstadt, Germany).

BAC Assay:

Article Title: Ginkgolic Acid is a Multi-Target Inhibitor of Key Enzymes in Pro-Inflammatory Lipid Mediator Biosynthesis
Article Snippet: .. Zileuton [N -(1-benzo[b]thien-2-ylethyl)-N -hydroxyurea] was from Sequoia Research Products (Oxford, UK); PGH2 was from Larodan (Malmö, Sweden); IL-1β was from ReproTech (Hamburg, Germany); RSC-3388 was from Calbiochem (Darmstadt, Germany); EDTA and Nonidet P-40 were from AppliChem (Darmstadt, Germany); p-anisidinium chloride was from Merck (Darmstadt, Germany); Insect Express Sf9-S2 and RPMI media, glutamine, penicillin, and streptomycin were from PAA (Coelbe, Germany); Bac-to-Bac baculovirus expression system was from Invitrogen (Karlsruhe, Germany); Ni-NTA agarose was from Qiagen (Hilden, Germany). .. AA, Ca2+ -ionophore A23187, dextrane, dithiothreitol, fetal calf serum, indomethacin, lipopolysaccharide (LPS), Triton X-100, and all other chemicals were purchased from Sigma-Aldrich (Taufkirchen, Germany), unless stated otherwise.

Marker:

Article Title: Oxidative status predicts quality in human mesenchymal stem cells
Article Snippet: Glycosaminoglycan (GAG) accumulation was determined as a chondrogenic marker. .. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l -cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem).

FACS:

Article Title: Binding of von Willebrand Factor to Complement C1q Decreases the Phagocytosis of Cholesterol Crystals and Subsequent IL-1 Secretion in Macrophages
Article Snippet: After 18 h, HMDMs were washed with PBS and incubated with PBS/10 mM EDTA (AppliChem, Darmstadt, Germany) for 30 min at 4°C. .. Cells were collected in FACS buffer (PBS/0.1% FCS/1 mM EDTA) and resuspended at a cell concentration of 5 × 105 cells/100 μl and incubated with 2 μg/ml of human IgG for 45 min at 4°C to block unspecific binding of antibodies to Fcγ receptors.

MTT Assay:

Article Title: Cellular Pharmacology of Palladinum(III) Hematoporphyrin IX Complexes: Solution Stability, Antineoplastic and Apoptogenic Activity, DNA Binding, and Processing of DNA-Adducts
Article Snippet: Agarose, ethanol, DMSO, formic acid, 2-propanol, methanol, EDTA, ethidium bromide, sodium chloride, Tris hydrochloride, Triton® X-100, l -glutamine were purchased from AppliChem GmbH, Darmstadt, Germany. .. The tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and TE buffer werе supplied by Merck, Darmstadt, Germany.

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