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Glyceraldehyde 3 phosphate dehydrogenase gapdh oligonucleotide primers

Manufactured by Bioneer

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers are short sequences of DNA designed to amplify a specific region of the GAPDH gene. GAPDH is a commonly used reference gene in gene expression analysis. These primers can be used in various molecular biology techniques, such as reverse transcription-polymerase chain reaction (RT-PCR), to quantify the expression levels of GAPDH.

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9 protocols using glyceraldehyde 3 phosphate dehydrogenase gapdh oligonucleotide primers

1

Adipocyte Differentiation Regulation

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3-Isobutyl-1-methylxanthine (IBMX), dexamethasone (DEX), insulin, and Oil Red O powder were purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Dulbecco's modified Eagle's medium (DMEM), bovine serum (BS), fetal bovine serum, and antibiotic-antimycotic (ABAM) were obtained from Life Technologies, Inc. (Grand Island, NY, USA). A 45% HFD was acquired from research diets (New Brunswick, NJ, USA). PPARγ, C/EBPα, SREBP1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer Corporation (Daejeon, Republic of Korea), and SYBR Premix Ex Taq was purchased from Takara Bio Inc. (Otsu, Japan). Antibodies against PPARγ (cat. No. sc-7273), C/EBPα (cat. No. sc-9314), SREBP1 (cat. No. sc-13551), and β-actin (cat. No. sc-81178) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Horseradish peroxidase-conjugated secondary antibodies were obtained from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).
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2

Adipocyte Differentiation Protocol

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BV, IBMX, Dexamethasone (DEX), insulin, Oil red O, and all other chemicals were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco’s modified Eagles medium (DMEM), bovine serum (BS), fetal bovine serum (FBS), and penicillin-streptomycin (PS) were purchased from Life Technologies, Inc. (Grand Island, NY, USA). Antibodies against PPARγ (E-8; cat. no. sc-7273), C/EBPα (C-18; cat. no. sc-9314), and β-actin (C4; cat. no. sc-47778) were purchased from Santa Cruz biotechnology, Inc. (Santa Cruz, CA, USA). Phospho-extracellular signal-regulated kinase (p-ERK; Thr202/Tyr204; cat. no. #9101), ERK (cat. no. #9102), phospho-stress-activated protein kinase/Jun-amino-terminal kinase (p-JNK; Thr183/Tyr185; cat. no. #9251), JNK (cat. no. #9252), phospho-p38 MAPK (p-p38; Thr180/Thy182; cat. no. #9215), p38 (cat. no. #9212), p-AMPK (Thr172; cat. no. #2535), AMPK (cat. no. #2532), p-ACC (Ser79; cat. no. #3661), and ACC (cat. no. #3662) antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA) Horseradish peroxidase conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA). SYBR Green Master Mix was purchased from Applied Biosystems (Foster, CA, USA). C/EBPα, C/EBPβ, C/EBPδ, PPARγ, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer (Daejeon, Korea).
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3

Adipogenic Effects of IBMX, DEX, and Insulin

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3-Isobutyl-1-methylxanthine (IBMX), dexamethasone (DEX), insulin, and Oil Red O powder were purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Dulbecco's modified Eagle's medium (DMEM), bovine serum (BS), fetal bovine serum (FBS), and Antibiotic-Antimycotic (ABAM) were purchased from Life Technologies, Inc. (Grand Island, NY, USA). Orlistat was purchased from Tokyo Chemical Inc. (Tokyo, Japan). HFD (30%) was obtained from Research Diets (New Brunswick, NJ, USA). PPARγ, C/EBPα, SREBP1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer Corporation (Daejeon, Republic of Korea), and SYBR Premix Ex Taq was purchased from Takara Bio Inc. (Otsu, Japan). Antibodies against PPARγ (cat. No. sc-7273), C/EBPα (cat. No. sc-9314), SREBP1 (cat. No. sc-13551), C/EBPβ (cat. No. sc-150), and β-actin (cat. No. sc-81178) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Horseradish peroxidase conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).
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4

HepG2 Cell Culture and Characterization

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The HepG2 human
hepatocellular liver cell line was obtained from the Korea Research
Institute of Bioscience and Biotechnology (Daejeon, South Korea) and
grown in Eagle’s minimum essential medium (EMEM) containing
10% fetal bovine serum and 100 U/mL penicillin/streptomycin sulfate.
Cells were incubated in a humidified 5% CO2 atmosphere
at 37 °C. EMEM, penicillin, and streptomycin were purchased from
Hyclone (Logan, UT, USA). Bovine serum albumin (BSA) and 25-hydroxycholesterol
were purchased from Sigma-Aldrich (St. Louis, MO, USA). 4′,6-Diamidino-2-phe-nylindole
(DAPI) and DiI-LDL were purchased from Thermo Fisher Scientific (Waltham,
MA, USA). Antibodies against LSS, SREBP1, SREBP2, HMGCR, SQLE, and
β-actin were purchased from Abcam, Inc. (Cambridge, MA, USA).
Antibodies against PCSK9 were purchased from Cell Signaling Technology.
(Beverly, MA, USA). SREBF1, SREBF2, HMGCR, PCSK9, IDI1, SQLE, ABCA1,
ACSL6, DHCR7, FDFT1, FDPS, SOAT1, and glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) oligonucleotide primers were purchased from Bioneer Corp.
(Daejeon, South Korea). α-Mangostin was isolated from the chloroform
fraction of G. mangostana, as previously
described, and confirmed by a spectroscopic analysis. The purity of
α-mangostin was determined to be over 95% by high-performance
liquid chromatography using an ultraviolet detector.4 (link)
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5

Herbal Extract Composition and Anti-Obesity Effects

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TBS consists of C. rotundus L., C. unshiu Markovich, and Poria cocos. The three herbs were purchased from Nanum Pharmaceutical Company (Seoul, Republic of Korea). The herbal samples were performed for sensory test according to ‘The Korean Herbal Pharmacopoeia’ by Prof. Yun-Yeop Cha, and only those that passed the Korean Pharmacopoeia standard were selected and used for this experiment. TBS was made using a 1: 1: 1 ratio of these herbs (400 g each). The herbs were then extracted in water at 99 °C for 3 h. The extract was freeze-dried, and the yield rate was calculated at 33.20% (33.20 g per 100 g of liquid extract). The powder was dissolved in distilled water for this experiment, and the residual powder was stored at − 20 °C. The 30% HFD was obtained from Research Diets (New Brunswick, NJ, USA). The p-AMPK and AMPK antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). PPARγ, C/EBPα, SREBP1, AMPK, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer Corporation (Daejeon, Republic of Korea), and SYBR Premix Ex Taq was purchased from Takara Bio Inc. (Otsu, Japan). Orlistat was purchased Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan) and other reagents were purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA).
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6

Molecular Mechanisms of Prostate Cancer Progression

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Testosterone propionate (Cat# 200-17532) was purchased from Wako Pure Chemicals (Tokyo, Japan). Finasteride was procured from Merck & Co., Inc. (NJ, USA). Antibodies against Bcl-2 (Cat# sc-7382), Bcl-xL (Cat# sc-8392), CDK2 (Cat# sc-748), CDK4 (Cat# sc-23896), Cyclin A (Cat# sc-751), Cyclin D1 (Cat# sc-753), Cyclin E (Cat# sc-481), E2F1 (Cat# sc-193), PCNA (Cat# sc-56), p-RB (Cat# sc-377528), and β-actin (Cat# sc-81178) were purchased from Santa Cruz biotechnology, Inc. (Dallas, TX, USA). Antibody against PSA (Cat# PB9259) was procured from Boster Biological Technology (Pleasanton, CA, USA). Horseradish peroxidase conjugated secondary antibodies (Cat# 315-035-003 and 111-035-003) were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA). SYBR Green Master Mix (Cat# 4367659) was purchased from Applied Biosystems (Foster City, CA, USA). 5α-reductase 2, CDK2, CDK4, Cyclin A, Cyclin D1, Cyclin E, E2F1, PCNA, PSA, RB, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer (Daejeon, Korea).
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7

HepG2 Cell Culture and Analysis

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The HepG2 human hepatocellular liver cell line was obtained from the Korea Research Institute of Bioscience and Biotechnology (Daejeon, Korea) and grown in Eagle’s Minimum Essential Medium (EMEM) containing 10% fetal bovine serum and 100 U/mL penicillin/streptomycin sulfate. Cells were incubated in a humidified 5% CO2 atmosphere at 37 °C. EMEM, penicillin, and streptomycin were purchased from Hyclone (Logan, UT, USA). Bovine serum albumin was purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against PCSK9, LDL-R, and β-actin were purchased from Abcam, Inc. (Cambridge, MA, USA). PCSK9, LDL-R, SREBP1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer Corp. (Daejeon, Korea). The solvents for extraction and isolation (methanol, ethyl acetate, n-butyl alcohol, chloroform, n-hexane, etc.) were purchased from SK Chemical (Seoul, Korea). The solvents for HPLC-grade acetonitrile (MeCN) and methanol were also purchased from SK Chemical. The solvent for NMR (CD3OD) was obtained from Cambridge Isotope Laboratories, Inc. (Andover, MA, USA).
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8

Cordycepin Modulates Inflammatory Pathways

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Cordycepin (MW, 251.2; Product No. C3394) from C. militaris, LPS (Escherichia coli 026:B6), Griess reagent, Tween-20, bovine serum albumin (BSA), H89, forskolin, compound C, and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Antibodies against COX-2, MMP-2, MMP-9, AMPK, pAMPK, ACC, pACC, CREB, pCREB, and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The peroxidase-labeled donkey anti-rabbit immunoglobulin, peroxidase-labeled sheep anti-mouse immunoglobulin, and enhanced chemiluminescence (ECL) detection kit were purchased from Amersham Corp. (Arlington Heights, IL, USA). The enzyme-linked immunosorbent assay (ELISA) kit for PGE2 was obtained from R&D Systems (Minneapolis, MN, USA). COX-2, EP1-4, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer (Seoul, Korea). All other chemicals were purchased from Sigma-Aldrich Chemical Co.
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9

Molecular Mechanisms of Testosterone Regulation

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Testosterone, phenylmethylsulfonyl fluoride, Triton-X-100 and the protein inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Finasteride, a type II 5-reductase inhibitor, was obtained from Merck & Co., Inc. (Whitehouse Station, NJ, USA). 5α-Reductase 2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer Corporation (Daejeon, Republic of Korea), and SYBR Premix Ex Taq was purchased from Takara Bio., Inc. (Otsu, Japan). Antibodies against inducible nitric oxide synthase (iNOS; M-19; cat. no. sc-650), cyclooxygenase 2 (COX-2; C-20; cat. no. sc-1745), poly (ADP-ribose) polymerase-1 (PARP-1; F-2; cat. no. sc-25780), caspase-3 (E-8; cat. no. sc-7272), Bcl-2 (C-2; cat. no. sc-7382), Bcl-xL (H-5; cat. no. sc-8392), Bax (B-9; cat. no. sc-7480) and β-actin (ACTBD11B7; cat. no. sc-81178) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). An antibody for proliferating cell nuclear antigen (PCNA; cat. no. sc-56) was purchased from BD Biosciences (San Jose, CA, USA). Horseradish peroxidase-conjugated secondary antibody was purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).
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