Immunoprecipitation:Article Title: The Regulation of COX-2, ABCA1 and ABCG1 by the lncRNA PACERR links the inflammatory response and cholesterol homeostasis
Article Snippet: In each experiment, chromatin was first processed by agarose gel electrophoresis to confirm DNA shearing to 200 - 500 bp fragments, and the DNA concentration was measured by NanoDrop 2000. .. Equal quantities of sheared chromatin (10 μg per immunoprecipitation) were diluted 1:5 in sonication buffer to the final volume of 1 mL, and immunoprecipitated overnight with 1 μg antibody targeting human hnRNPL (novus NB120-6106), P300 (Abcam ab275378), Histone H3K27Ac (Active Motif, 39685), IkBζ (Cell Signaling 9244S), cPLA2 (Cell Signaling 2832S) and LXR-α and LXR-β (Active motif 61175 and 61177). ..
Sonication:Article Title: The Regulation of COX-2, ABCA1 and ABCG1 by the lncRNA PACERR links the inflammatory response and cholesterol homeostasis
Article Snippet: In each experiment, chromatin was first processed by agarose gel electrophoresis to confirm DNA shearing to 200 - 500 bp fragments, and the DNA concentration was measured by NanoDrop 2000. .. Equal quantities of sheared chromatin (10 μg per immunoprecipitation) were diluted 1:5 in sonication buffer to the final volume of 1 mL, and immunoprecipitated overnight with 1 μg antibody targeting human hnRNPL (novus NB120-6106), P300 (Abcam ab275378), Histone H3K27Ac (Active Motif, 39685), IkBζ (Cell Signaling 9244S), cPLA2 (Cell Signaling 2832S) and LXR-α and LXR-β (Active motif 61175 and 61177). ..
Protease Inhibitor:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Transfection:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Saline:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Radio Immunoprecipitation:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Bicinchoninic Acid Protein Assay:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Western Blot:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
Stripping Membranes:Article Title: Cytosolic phospholipase A2 links tau pathology to insulin signaling impairment in Alzheimer’s disease
Article Snippet: .. Tau-441 from rPeptide (Bogart, GA), and primary antibodies and reagents were purchased as follows: Anti-tau (T22) from Millipore Sigma (St. Louis, MO); antibodies against IR-α, IR-β, IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Tyr895), cPLA2, p-cPLA2 and Cav-1 from Cell Signaling (Beverly, MA); dimethyl sulfoxide (DMSO), cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor cocktail, β-mercaptoethanol (β-ME), Lipofectamine 3000 transfection reagent and cPLA2 siRNA from Thermo Fisher Scientific (Waltham, MA); Ham’s F-12 Nutrient Mix from Crystalgen Inc., (Commack, NY); EGM-2 MV BulletKit from Lonza (Walkersville, MD); penicillin/streptomycin (P/S), Hanks’ Balanced Salt Solution (HBSS), and Dulbecco’s phosphate-buffered saline (DPBS) from Life Technologies (Grand Island, NY); fibronectin, collagen from calf skin, laminin, and Minimum Essential Media Eagle (MEM) from Millipore Sigma (St. Louis, MO); papain and DNase from Worthington Biochemical (Lakewood, NJ); radio-immunoprecipitation assay (RIPA) buffer, BCA protein assay kit, SuperSignalTM West Femto maximum sensitivity substrate, SuperSignalTM West Pico Plus chemiluminescent substrate, and RestoreTM PLUS western-blot stripping buffer from Thermo Fisher Scientific (Waltham, MA); and Laemmli sample buffer and tris-buffered saline (TBS) from Bio-Rad (Hercules, CA). ..
other:Article Title: Cytosolic phospholipase A2 as a therapeutic target for degenerative joint diseases
Article Snippet: To assess the therapeutic effects of fexofenadine (FFD) or AACOCF3 (a known cPLA2 inhibitor) across different genetic backgrounds, mice received oral gavage of either vehicle (5% DMSO + 10% ethanol), AACOCF3 (10 mg/kg, sc-201412A, Santa Cruz), or FFD (10 mg/kg, S3208, Selleckchem) for 8 weeks, starting 28 days post-DMM surgery.
Incubation:Article Title: Cytosolic phospholipase A2 as a therapeutic target for degenerative joint diseases.
Article Snippet: For IHC, deparaffinized and hydrated sections were incubated with 0.1% trypsin for 30minutes at 37 °C, followed by treatment with 0.25 U/mL chondroitinase ABC (Sigma) and 1 U/mL hyaluronidase (Sigma) for 60min at 37 °C. .. Sections were incubated overnight at 4 °C with primary antibodies against Col II (ProteinTech, Cat. 28459-1-AP), aggrecan neoepitope (Novus Biologicals, Cat. NB100-74350), MMP13 (ProteinTech, Cat. 18165-1-AP), cPLA2 (CST, Cat. 2831), p16 (CST, Cat. 18769), and p21 (Invitrogen, Cat. MA514949). .. Detection was performed with the Vectastain Elite ABC kit (Vector Laboratories, PK-6100), and positive signals were visualized using 0.5 mg/mL 3,3-diaminobenzidine in a 50mmol/L Tris-HCl substrate (Sigma).
Article Title: Cytosolic phospholipase A2 as a therapeutic target for degenerative joint diseases
Article Snippet: For IHC, deparaffinized and hydrated sections were incubated with 0.1% trypsin for 30 minutes at 37 °C, followed by treatment with 0.25 U/mL chondroitinase ABC (Sigma) and 1 U/mL hyaluronidase (Sigma) for 60 min at 37 °C. .. Sections were incubated overnight at 4 °C with primary antibodies against Col II (ProteinTech, Cat. 28459-1-AP), aggrecan neoepitope (Novus Biologicals, Cat. NB100-74350), MMP13 (ProteinTech, Cat. 18165-1-AP), cPLA2 (CST, Cat. 2831), p16 (CST, Cat. 18769), and p21 (Invitrogen, Cat. MA5-14949). .. Detection was performed with the Vectastain Elite ABC kit (Vector Laboratories, PK-6100), and positive signals were visualized using 0.5 mg/mL 3,3-diaminobenzidine in a 50 mmol/L Tris-HCl substrate (Sigma).
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