Celastrol was purchased from Calbiochem (San Diego, CA, USA) and dissolved in DMSO. Cobalt chloride, N-acetylcysteine (NAC) and cycloheximide (CHX) were purchased from Sigma (St Louis, MO, USA). LY294002 was purchased from Alexis (San Diego, CA, USA). The antibodies used were as follows: anti-HIF-1α and anti-HIF-1β (BD Transduction Laboratories, San Jose, CA, USA); anti-Raf, anti-Akt, anti-p-Akt (S473), anti-p-p70S6K (T389) and anti-PARP (Cell Signaling, Beverly, MA, USA); anti-BNIP3, anti-p53 and anti-p21 (Santa Cruz, Dallas, TX, USA); and anti-Bcl2, anti Bcl-xL and anti-β-actin (Calbiochem, San Diego, CA, USA). The dual-Luciferase reporter assay system was purchased from Promega (Madison, WI, USA).
Anti bnip3
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-BNIP3 is a primary antibody that specifically recognizes the BNIP3 protein. BNIP3 is a pro-apoptotic member of the Bcl-2 family and plays a role in mitochondrial-dependent cell death pathways.
Automatically generated - may contain errors
Lab products found in correlation
Anti hif 1α, by BD (2 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Anti pakt s473, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Anti p p70s6k t389, by Cell Signaling Technology (1 mentions)
Cobalt chloride, by Merck Group (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
Anti hif 1β, by BD (1 mentions)
Anti raf, by Cell Signaling Technology (1 mentions)
Anti p21, by Santa Cruz Biotechnology (1 mentions)
Anti p53, by Santa Cruz Biotechnology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Anti lat, by Cell Signaling Technology (1 mentions)
Anti myc tag, by Cell Signaling Technology (1 mentions)
Anti β actin, by BD (1 mentions)
Anti lamin b, by Santa Cruz Biotechnology (1 mentions)
Anti hmgcr, by Santa Cruz Biotechnology (1 mentions)
Anti p yap, by Cell Signaling Technology (1 mentions)
Anti p lats, by Cell Signaling Technology (1 mentions)
4 protocols using anti bnip3
1
Celastrol Modulates Hypoxia Signaling
2
Immunoblotting Protein Analysis in Cells
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Immunobloting analysis of proteins in cell lysates and nuclear fractions was performed as previously described [56 (link), 57 (link)]. Primary antibodies used were as follows: anti-YAP (#4912), anti-p-YAP (#4911), anti-Myc-Tag (#2278), anti-Mst (#3682), anti-p-Mst (#3681), anti-LATS (#3477) and anti-p-LATS (#8654) were purchased from Cell Signaling Technology (Beverly, MA); anti-HIF-1α (#610959) was purchased from BD; anti-β-Actin (#Sc-47778), anti-Lamin B (#Sc-6216), anti-BNIP3 (#Sc-56167), and anti-HMGCR (#27578) were purchased from Santa Cruz (Santa Cruz, CA).
3
Investigating IGF-1 Signaling in C2C12 Myoblasts
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C2C12 myoblasts were purchased from ATCC (Shanghai, China). Dulbecco modified Eagle’s medium (DMEM), penicillin/streptomycin (P/S), fetal bovine serum (FBS), and horse serum (HS) were purchased from Thermo Scientific (Waltham, MA, USA). IGF-1, BMS754807 (BMS), 3-MA, and chloroquine (CQ) were purchased from MedChemExpress (Shanghai, China). Antibodies such as anti-phospho-IGF-1R, anti-IGF-1R, anti-phospho-AKT, anti-AKT, anti-phospho-mTOR, and anti-mTOR were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-BNIP3, anti-myosin heavy chain (MHC), anti-Myogenin (MyoG), and anti-TOMM20 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies such as anti-α-tubulin, anti-GAPDH, anti-PGC-1α, anti-p62, and anti-LC3 were obtained from Protientech (Wuhan, China).
4
Western Blot Analysis of Cellular and Exosomal Proteins
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Total cell lysates (20 μg) or exosome lysates (15 μg) were separated on 10% SDS-PAGE and transferred to a polyvinylidene difluoride membrane (Millipore). After blocking, the membranes were blotted with the following antibodies: anti-HSP70 (Cat. No. sc33575), anti-CD63 (Cat. No. sc15363), anti-NPM1 (Cat. No. sc4772) (Santa Cruz, Dallas, TX), anti-BNIP3 (Cat. No. ab10433), anti-CD9 (Cat. No. ab92726) (Abcam, Cambridge, UK), anti-β-actin (Cat. No. #4970), anti-CCNB1 (Cat. No. #4135), anti-MRE11 (Cat. No. #4895S), anti-HMGB1 (Cat. No #3935), anti-RhoA (Cat. No. #2177) (Cell Signaling Technology, Danvers, MA, USA) at a dilution of 1:1000, as well as anti-HGS (Cat. No. WH0009146), anti-KHSRP (Cat. No. HPA034739) and anti-PES1 (Cat. No. HPA062439) (Sigma-Aldrich) at a dilution of 1:500. Following intensive washing, the membranes were developed with anti-mouse or rabbit horseradish peroxidase conjugated secondary antibodies (Cell Signaling Technology) at a dilution of 1:5000. The protein bands were visualized using Super Signal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, Waltham, MA, USA) with the ImageQuant LAS4000 system (Fujifilm, Tokyo, Japan).
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