Biib021

Manufactured by Selleck Chemicals

Sourced in United States

BIIB021 is a laboratory instrument designed for the analysis and characterization of biomolecules, such as proteins and nucleic acids. It utilizes advanced techniques to provide detailed information about the structure and properties of these important biological molecules.

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9 protocols using biib021

Modulating Protein Aggregation in Fibroblasts

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For activation of the HS pathway in human and mouse fibroblasts, an established HSP90 inhibitor, BIIB021 (Selleckchem, Houston, TX), was used at 100 nM concentration (Lundgren et al., 2009 (link)). To inhibit the proteasome, Wt and HSP70.1/3^−/− MEFs were plated in 2% FCS and allowed to proliferate for 24 hr. Following this, the cultures were treated with 30 μM lactacystin (Lc) (Biomol Research Laboratories, PA) or its vehicle dimethyl sulfoxide (DMSO; Sigma, St. Louis, MO), for 16 hr in 0% FCS (Notterpek et al., 1999 (link)). Treatment conditions were selected after testing multiple doses and paradigms to achieve aggregate count in 50% of the cells (Fortun et al., 2007 (link)). This experimental model allowed us to assess the effects of BIIB021 and 3-methyladenine (3-MA; Sigma, St. Louis, MO) on aggregate formation. The compound 3-MA is a known inhibitor of autophagy through class III phosphatidylinositol 3-kinases when used at 4 mM for 16 hr (Wu et al., 2010 (link)).

Chittoor-Vinod V.G., Lee S., Judge S.M, & Notterpek L. (2015). Inducible HSP70 Is Critical in Preventing the Aggregation and Enhancing the Processing of PMP22. ASN NEURO, 7(1), 1759091415569909.

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Cell Line Characterization and Drug Treatments

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Ramos, Raji, Namalwa, Daudi, CA-46, Ly10 and Ly18 cell lines were purchased from American Type Culture Collection (ATCC) in 2013. Jiyoye and DG-75 were purchased from ATCC in 2014. Raji 2R and Raji 4RH were provided by Roswell Park Cancer Institute (M.J.B.). All cells were used for experiments within 2 months of thawing. Cell line characterization was performed by Idex Bioresearch (Westbrook, Maine) using short tandem repeat profiling and multiplex PCR to report cell line characterization and interspecies contamination. Testing was last performed in February 2014. Cells were cultured in RPMI-1640 media (Invitrogen) supplemented with 10% heat inactivated fetal bovine serum and gentamicin 50ug/mL (Sigma-Aldrich). PU-H71, PU-H71-beads, and control beads were synthesized and prepared at Memorial Sloan Kettering Cancer Center as previously reported(13 (link),14 (link)). Idelalisib (CAL-101), IPI-145, BKM120 (15 (link)), BEZ-235 (16 (link)), ganetespib (17 (link)), and BIIB-021 (18 (link)) were purchased from Selleck Chemicals. MPC-3100 (19 (link)) and CUDC-305 (20 (link)) were purchased from Santa Cruz and ChemieTek respectively. The chemical structures for PU-H71 and BEZ-235 are presented in Supplemental Figure 1.

Giulino-Roth L., van Besien H.J., Dalton T., Totonchy J.E., Rodina A., Taldone T., Bolaender A., Erdjument-Bromage H., Sadek J., Chadburn A., Barth M.J., Dela Cruz F.S., Rainey A., Kung A.L., Chiosis G, & Cesarman E. (2017). Inhibition of Hsp90 suppresses PI3K/AKT/mTOR signaling and has antitumor activity in Burkitt lymphoma. Molecular cancer therapeutics, 16(9), 1779-1790.

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Evaluating HSP90 Inhibitors in Schwann and DRG Cells

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HSP90 inhibitor
compounds, including AT13387 (S1163), AUY922 (S1069), BIIB021 (S1175),
SNX5422 (S2656), and STA9090 (S1159), were purchased from Selleckchem
(Houston, TX) and stored at a stock concentration of 1 mM in DMSO.
Primary Schwann cells were treated with HSP90 inhibitors at the indicated
concentrations in complete media (see above), 24 h after seeding.
DMSO served as the vehicle control while geldanamycin (GA) was used
as a positive control for heat shock pathway activation. The DRG explant
cultures were maintained for 7 days in ascorbate-containing media
prior to treatment with either DMSO, AUY922 (100 nM), or BIIB021 (100
nM), every third day (72 h apart). Cultures were procured, 24 h after
the third treatment, for either biochemical or immunochemical analyses.^{15 (link)}

Chittoor-Vinod V.G., Bazick H., Todd A.G., Falk D., Morelli K.H., Burgess R.W., Foster T.C, & Notterpek L. (2019). HSP90 Inhibitor, NVP-AUY922, Improves Myelination in Vitro and Supports the Maintenance of Myelinated Axons in Neuropathic Mice. ACS Chemical Neuroscience, 10(6), 2890-2902.

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Melanoma Cell Culture and Inhibitors

Cited 2 times

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Culture conditions and the cell lines have been previously described [12] (link). The B16-BL6 murine melanoma cell line was provided by Dr. Andrew Hurwitz [24] (link). Human cell lines were obtained from the ATCC, (Manassas, VA) (including MALME-M3, MM96L, A375, 453A, MM455, Roth, H59-44T, Mel-Juso, JURKAT), or from a patient at the Massashusetts General Hospital (MU89 and MU-X (derived from MU89 as previously described [6] (link), [11] (link)). Construction and culture of the J.RT3-T3.5 cell line expressing a Melan-A/MART-1 specific TCR has been described [15] (link). IFN-β-1a (Avonex) was obtained from Biogen-Idec (Cambridge, MA). The iHsp90s were obtained from the following sources: Radicicol (A.G. Scientific, San Diego, CA); Novobiocin (BioMol, Plymouth Meeting, PA); 17-DMAG (LC laboratories, Woburn, MA); 17-AEP (InVivoGen, San Diego, CA); Rifabutin, PU-H71, and 17-AAG (Sigma, St. Louis, MO); Gedunin, CCT018159, and celastrol (Tocris, Ellisville, MO); and NVP-AUY922 and BIIB021 (Selleck Chemicals, Houston, TX).

Haggerty T.J., Dunn I.S., Rose L.B., Newton E.E., Pandolfi F, & Kurnick J.T. (2014). Heat Shock Protein-90 Inhibitors Enhance Antigen Expression on Melanomas and Increase T Cell Recognition of Tumor Cells. PLoS ONE, 9(12), e114506.

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Pressure-Induced NPSC Response Modulation

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To mimic the in vivo compression environment, a stainless-steel pressure vessel was utilized following the protocols previously established by our group (Chen et al., 2017 (link), 2018 (link); Li et al., 2018a (link); Wang W. et al., 2018 (link)). NPSCs which were cultured in cell culture dishes or plates as monolayer were placed on the bracket in the pressure apparatus and exposed to 1.0 MPa pressure for different time periods. The pressure vessel was supplied with a small quantity of double distilled water at the bottom to maintain moisture and placed in an incubator at 37°C.
The HSP90 inhibitor BIIB021, RIPK1 inhibitor Necrostatin-1 (Nec-1), RIPK3 inhibitor GSK′872, MLKL inhibitor Necrosulfonamide (NSA), proteasome inhibitor MG132, and HSP70 inhibitor Ver155008 (Ver) were all purchased from Selleck Chemicals (Houston, TX, United States). For all experiments, NPSCs were pretreated with inhibitors for 2 h before compression treatment, while control groups were given isopyknic dimethylsulfoxide as vehicle control.

Hu B., Zhang S., Liu W., Wang P., Chen S., Lv X., Shi D., Ma K., Wang B., Wu Y, & Shao Z. (2020). Inhibiting Heat Shock Protein 90 Protects Nucleus Pulposus-Derived Stem/Progenitor Cells From Compression-Induced Necroptosis and Apoptosis. Frontiers in Cell and Developmental Biology, 8, 685.

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Evaluation of Small Molecule Inhibitors

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The drugs employed in this study were as follows: dabrafenib (ApexBio, B1407‐50); XL888 (ApexBio, A4388‐25); fludarabine (Selleckchem, S1491); CHIR‐99021 HCl (CT99021) (Selleckchem, S2924); palbociclib (ApexBio, A8316); LDC000067 (ApexBio, B4754‐10); Ro 3306 (ApexBio, A8885‐10); roscovitine (ApexBio, A1723‐10); K03861 (Selleckchem, S8100); CHIR‐99021 (Selleckchem, S2924); dinaciclib (Selleckchem, S2768); FRAX597 (Selleckchem, S7271); PF‐3758309 (Selleckchem, S7094); AUY922 (Selleckchem, S1069); BIIB021 (Selleckchem, S1175); novobiocin (Selleckchem, S2492); 17‐DMAG (Selleckchem, S1142). All the drugs were dissolved in DMSO (Sigma D2650).

Azimi A., Caramuta S., Seashore‐Ludlow B., Boström J., Robinson J.L., Edfors F., Tuominen R., Kemper K., Krijgsman O., Peeper D.S., Nielsen J., Hansson J., Egyhazi Brage S., Altun M., Uhlen M, & Maddalo G. (2018). Targeting CDK2 overcomes melanoma resistance against BRAF and Hsp90 inhibitors. Molecular Systems Biology, 14(3), e7858.

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Comprehensive Molecular Profiling Assay

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BIIB021 (#S1175) and TAK-733 (#S2617) were obtained from Selleck
Chemicals (Houston, TX). Bardoxolone methyl (#1772) was obtained from Axon
MedChem (Groningen, Netherlands). PD0325901 (P-9688) and GDC-0941 (G-9252) was
obtained from LC Laboratories (Woburn, MA). Marizomib (SalA-100) was obtained
from FIVEphoton Biochemicals (San Diego, CA). Neurocult NS-A Proliferation kit
(#05751) and heparin (#07980) were obtained from StemCell Technologies
(Vancouver, British Columbia). Basic-FGF (AF-100–18) and EGF
(AF-100–15) were obtained from PeproTech (Rocky Hill, NJ). alamarBlue
(DAL1100) was obtained from ThermoFisher Scientific (Waltham, MA). CellTiter-Glo
(G7573) and Caspase-Glo 3/7 (G8092) were obtained from Promega (Madison, WI).
NF-1 antibody (sc-67) and beta-actin antibody (sc-47778 HRP) were obtained from
Santa Cruz Biotechnology (sc-67). PTEN antibody (#9559T) and goat anti-rabbit
IgG (7074P2) secondary antibody were obtained from Cell Signaling Technology
(Danvers, MA). Phosphokinase antibody array (ARY003B) was obtained from R&D
Systems (Minneapolis, MN).

Wilson K.M., Mathews-Griner L.A., Williamson T., Guha R., Chen L., Shinn P., McKnight C., Michael S., Klumpp-Thomas C., Binder Z.A., Ferrer M., Gallia G.L., Thomas C.J, & Riggins G.J. (2018). Mutation Profiles in Glioblastoma 3D Oncospheres Modulate Drug Efficacy. SLAS technology, 24(1), 28-40.

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Evaluation of BIIB021 Cytotoxicity in SKM-1 Cells

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SKM-1 cells (JCRB0118; Japanese Collection of Research Bioresources Cell Bank, Osaka, Japan) were cultured in RPMI-1640 medium (Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (Gibco) at 37°C in a humidified atmosphere of 5% CO₂. BIIB021 was purchased from Selleck Chemicals (Houston, TX, USA). Methylcellulose and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All antibodies used in the western blot analysis were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), with the exception of human anti-β-actin. Insulin-like growth factor-1 (IGF-1) was purchased from Peprotech (Rocky Hill, NJ, USA). The caspase inhibitors z-IETD-fmk and z-LEHD-fmk were purchased from Biovision (Palo Alto, CA, USA).

LIN S., LI J., ZHOU W., QIAN W., WANG B, & CHEN Z. (2014). BIIB021, an Hsp90 inhibitor, effectively kills a myelodysplastic syndrome cell line via the activation of caspases and inhibition of PI3K/Akt and NF-κB pathway proteins. Experimental and Therapeutic Medicine, 7(6), 1539-1544.

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Screening of HSP90 inhibitor compounds

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The following drugs were purchased from Selleck Chem: 17-AAG (Cat No: S1141), NMS-E973 (Cat No: S7282), Onalespib (Cat No: S1163), XL888 (Cat No: S7122), Alvespimycin (17-DMAG) HCl (Cat No: S1142), VER-49009 (Cat No: S7458), BIIB021 (Cat No: S1175), SNX-2112 (Cat No: S2639), and Anisomycin (S7409). Lysophosphatidic acid (LPA, L7260) was purchased from Sigma. All drugs were pre-arrayed into a 384-well source plate and serial dilutions were made in 100% DMSO. A fixed 100-nL volume was then transferred from the source plate to assay wells using a pin tool (Tecan). Screens were performed at 10 doses with two technical (on-plate) replicates. Each assay plate contained 16 DMSO-treated negative controls, 16 non-treated controls, and 16 Background (No Cells) controls.

Powell R.T., Moussalli M.J., Guo L., Bae G., Singh P., Stephan C., Shureiqi I, & Davies P.J. (2022). deepOrganoid: A brightfield cell viability model for screening matrix-embedded organoids. SLAS discovery : advancing life sciences R & D, 27(3).

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