The sample of Bruceine D (≥98% purity) used in the current study was provided by the Institute of Traditional Chinese Medicine and Natural Products, Jinan University (Guangzhou, China). RPMI-1640 medium and FBS were purchased from Gibco; Thermo Fisher Scientific, Inc. The MTT cell proliferation and cytotoxicity assay kits and penicillin/streptomycin solution (PS) were purchased from Nanjing KeyGen Biotech Co., Ltd. The antibody targeting PI3K (cat. no. AF5112) was obtained from Affinity Biosciences. Antibodies against AKT (cat. no. 60203-2-Ig), phosphorylated (p)-AKT (cat. no. 66444-1-Ig), E-cadherin (cat. no. 20874-1-AP) and β-catenin (cat. no. 51067-2-AP) were purchased from Proteintech Group, Inc. Vimentin antibody (cat. no. BS1491) was purchased from Bioworld Technology, Inc. Antibodies against GAPDH (cat. no. ab181602) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit (cat. no. ab6721) immunoglobulin (Ig) G were purchased from Abcam.
Immunoglobulin ig g
Manufactured by Abcam
Immunoglobulin (Ig) G is a type of antibody found in the blood and body fluids. It is the most abundant antibody in the human body and plays a critical role in the adaptive immune response. IgG helps neutralize pathogens, activate the complement system, and mark infected or foreign cells for destruction by other immune cells.
Automatically generated - may contain errors
Lab products found in correlation
Ez magna rip kit, by Merck Group (2 mentions)
Vimentin antibody, by Proteintech (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Ab133734, by Abcam (1 mentions)
Ab51266, by Abcam (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Anti fus antibody, by Abcam (1 mentions)
3 protocols using immunoglobulin ig g
1
Bruceine D Inhibits Cancer Progression
2
RNA Immunoprecipitation Assay for Protein-RNA Interactions
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The RNA immunoprecipitation (RIP) assay was conducted by using a commercial EZ‐Magna RIP™ Kit (Millipore) according to the manufacturer's instructions. In brief, cells (5 × 106) were treated with RIP lysis buffer at 4°C for 30 min and then whole‐cell lysates were incubated with RIP buffer containing magnetic beads conjugated to antibodies against Ago2 (Millipore) or anti‐PTBP1 antibody (ab133734; Abcam) or anti‐SNRP70 antibody (ab51266; Abcam) or immunoglobulin (Ig) G (Abcam) for 2 h at RT with gentle shaking. The coprecipitated RNAs associated with PTBP1 were extracted with the Trizol reagent (Life Technologies) and analysed by quantitative real‐time PCR (qPCR). Enrichment associated with SNRP70 and species‐matched normal IgG served as positive and negative RIP controls respectively. Total RNA was considered as input controls.
3
Ago2 and FUS RIP Assay
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An EZ-Magna RIP kit (Millipore, USA) was used for the RIP assay. Briefly, bEnd.3 cells (1 × 10 7 ) were lysed at 4°C for 30 min, and whole-cell lysates were then incubated with RIP buffer containing magnetic beads conjugated to antibodies against Ago2 (Millipore), anti-FUS antibody (Abcam), or immunoglobulin (Ig) G (Abcam) for 2 h. Finally, qPCR assays were performed to analyze the precipitated RNAs.
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