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Gp91ds tat

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Gp91ds-tat is a laboratory reagent produced by AnaSpec. It functions as a cell-permeable peptide inhibitor that can be used in research applications.

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Lab products found in correlation

Fluo 4 am, by Thermo Fisher Scientific (3 mentions) Apocynin, by Merck Group (3 mentions) Colchicine, by Merck Group (2 mentions) N acetyl cysteine (nac), by Merck Group (2 mentions) Gsk1016790a, by Merck Group (2 mentions) Fluo 4 am ester, by Thermo Fisher Scientific (2 mentions) Gsk2193874, by Merck Group (2 mentions) Rhod 2 am, by Thermo Fisher Scientific (2 mentions) N acetyl l cysteine nac, by Fujifilm (2 mentions) Monoclonal rat anti cd68 antibody clone fa 11, by Bio-Rad (2 mentions) Anti ly 6g antibody clone rea526, by Miltenyi Biotec (2 mentions) Diphenyleneiodonium chloride dpi, by Cayman Chemical (2 mentions) L leucyl l leucine methyl ester hydrochloride, by Cayman Chemical (2 mentions) Hoechst 33342, by Dojindo Laboratories (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Scramble ds tat, by AnaSpec (2 mentions) L name, by Merck Group (2 mentions) Mito tempo, by Santa Cruz Biotechnology (2 mentions) Infinite horizons impactor, by Precision Systems and Instrumentation (1 mentions) Scrambled ds tat, by AnaSpec (1 mentions)

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Gp91ds-tat peptide (3 citations)

27 protocols using gp91ds tat

1

Spinal Cord Injury and Antioxidant Treatment

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Adult male C57Bl6 mice were utilized in all experiments (20–25 g, Taconic Farms, Derwood, MD). Mice were group housed and received food and water ad libitum with a 12:12 h light cycle. A total of 111 male mice were used for this study; 13 mice were removed from the study due to post-surgical complications. All experiments complied fully with the principles set forth in the “Guide for the Care and Use of Laboratory Animals” and were approved by the Uniformed Services University IACUC.
All subjects undergoing surgery received isoflurane (Primal Healthcare, Andhra Pradesh, India). Mice received a laminectomy, followed by a contusion simulating moderate SCI using the Infinite Horizons Impactor (50 kdyn; Precision Systems and Instrumentation, Fairfax Station, VA). Mice were immediately given an intrathecal injection of either gp91ds-tat or scrambled ds-tat (AnaSpec, Inc., Fremont, CA) diluted to 50 μM in saline in a 5 μl volume at the lesion epicenter. After expelling the liquid, the needle was held under the dura for 30 s prior to removal. The incision was then closed, and animals were maintained on heating pads until mice regained movement. Acetaminophen (Children’s Tylenol, 200 mg/kg) was added to drinking water for 72 h post-injury. Manual bladder expression was performed daily until normal bladder expression returned. Naïve mice did not undergo surgery or receive isoflurane.
Khayrullina G., Bermudez S, & Byrnes K.R. (2015). Inhibition of NOX2 reduces locomotor impairment, inflammation, and oxidative stress after spinal cord injury. Journal of Neuroinflammation, 12, 172.
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2

Characterization of Metastatic Breast Cancer Cells

Cited 3 times
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Human breast MCF10A epithelial cells were obtained from the American Type Culture Collection. The creation of cells and tumorigenic capacity are described in ref. 38 (link), and metastatic capacity in ref. 39 (link). The creation of the PTEN−/− cells has been described previously (40 (link), 41 (link)). Reagents used include 4 µM Fluo-4 AM (Life Technologies; F14201), hydrogen peroxide (H2O2) (Sigma-Aldrich; 21673), N-acetylcysteine (NAC) (USP Reference Standard 1009005), GP91ds-TAT (Anaspec; AS-63818) (42 (link)), colchicine (Sigma-Aldrich; C9754), and RQ-00203078 (Tocris; 5388).
Pratt S.J., Lee R.M., Chang K.T., Hernández-Ochoa E.O., Annis D.A., Ory E.C., Thompson K.N., Bailey P.C., Mathias T.J., Ju J.A., Vitolo M.I., Schneider M.F., Stains J.P., Ward C.W, & Martin S.S. (2020). Mechanoactivation of NOX2-generated ROS elicits persistent TRPM8 Ca2+ signals that are inhibited by oncogenic KRas. Proceedings of the National Academy of Sciences of the United States of America, 117(42), 26008-26019.
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3

Fluorescence Imaging Assay Reagents

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Taxol, colchicine, GSK2193874, GSK1016790A, NAC, and PTL were purchased from Sigma. BAPTA AM ester was from Cayman Chemical. GP91ds-TAT was from AnaSpec. SiR-tubulin was from Cytoskeleton Inc. CellROX Deep Red Reagent and Fluo-4-AM ester were purchased from Thermo Fisher Scientific.
Lyons J.S., Joca H.C., Law R.A., Williams K.M., Kerr J.P., Shi G., Khairallah R.J., Martin S.S., Konstantopoulos K., Ward C.W, & Stains J.P. (2017). Microtubules tune mechanotransduction through NOX2 and TRPV4 to decrease sclerostin abundance in osteocytes. Science signaling, 10(506), eaan5748.
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4

Evaluating gp91ds-tat in Traumatic Brain Injury

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A selective NOX2 inhibitor, gp91ds-tat (5mg/kg), or equal concentration of ds-tat scrambled peptide (AnaSpec Inc. Fremont, CA) was delivered by intraperitoneal injection (I.P.) to WT CCI mice (n=8/group) starting at 1 day post-injury, with repeated I.P. administrations at 2 and 3 days post-injury. gp91ds-tat dosing was based on reported therapeutic effects in mice (Abais et al., 2013 (link)). ds-tat scrambled- and gp91ds-tat-treated CCI mice underwent motor function testing (beam walk) on 1, 3, 5, and 7 days post-injury, and cognitive function testing (Y-maze) at 6 days post-injury. Mice were anesthetized (100 mg/kg sodium pentobarbital, I.P.) at 7 days post-injury and transcardially perfused with ice-cold 0.9% saline (100ml), followed by 300 ml of 4% paraformaldehyde. Brains were removed and post-fixed in 4% paraformaldehyde overnight, and cryoprotected in 30% sucrose for histological analysis. A non-treated sham control group of WT mice (n=6) were used to establish baseline motor and cognitive function performance.
Kumar A., Barrett J.P., Alvarez-Croda D.M., Stoica B.A., Faden A.I, & Loane D.J. (2016). NOX2 drives M1-like microglial/macrophage activation and neurodegeneration following experimental traumatic brain injury. Brain, behavior, and immunity, 58, 291-309.
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5

Amyloid-beta Induced Oxidative Stress

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Aβ peptide (Aβ1-42) was from Bachem Inc. (Torrance, CA, USA). Fluo4-AM, MitoSOXTM Red Mitochondrial Superoxide Indicator, MitoTracker® Orange CMTMRos, anti-rabbit Alexa Fluor® 488 and anti-mouse Alexa Fluor® 635 were from Molecular Probes, Inc. (Eugene, OR, USA). Hexafluoro-2-propanol (HFIP) and CMC were from Merck (Darmstadt, Germany), Neurobasal and Dulbecco’s modified essential medium (DMEM), B27 supplement and lipofectamine 2000 were from Gibco (Carlsbad, CA, USA). DOTAP Liposomal Transfection Reagent was from Sigma–Aldrich (Oakville, ON, Canada). Phosphodiester oligonucleotides (ODNs) were from Integrated DNA Technologies (Coralville, IA, USA). The mito-Pericam plasmid was donated by Dr. V. Eisner. Bicinchoninic acid assay (BCA) kit and mHsp-70 antibody were from Pierce Biotechnology (Rockford, IL, USA). Ryanodine was from Alexis (Lausen, Switzerland). PDVF membranes were from Millipore (Bedford, MA, USA). RyR2 antibody and Rhod2-AM was from Thermo-Fisher (Waltham, MA, USA). Gp91 ds-tat was from AnaSpec (Fremont, CA, USA).
SanMartín C.D., Veloso P., Adasme T., Lobos P., Bruna B., Galaz J., García A., Hartel S., Hidalgo C, & Paula-Lima A.C. (2017). RyR2-Mediated Ca2+ Release and Mitochondrial ROS Generation Partake in the Synaptic Dysfunction Caused by Amyloid β Peptide Oligomers. Frontiers in Molecular Neuroscience, 10, 115.
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6

Vasodilation Reagent Preparation

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Gp91ds-tat was from Anaspec (Fremont, CA, USA), and sodium nitroprusside was from MP Biomedicals (Solon, OH, USA). All other drugs were from Sigma-Aldrich (St. Louis, MO, USA). Stock solutions were prepared according to the manufacturer’s instructions, and diluted in PSS to the required concentrations before use.
Meyer M.R., Rosemann T., Barton M, & Prossnitz E.R. (2017). GPER Mediates Functional Endothelial Aging in Renal Arteries. Pharmacology, 100(3-4), 188-193.
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7

VTA Cannulation and Oxidative Stress Modulation in Rats

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Sprague-Dawley rats were anesthetized with a mixture of air and isoflurane and placed in a stereotaxic frame. Cannula (Plastic One) was implanted unilaterally into VTA [coordinates: AP +3.4 (Lambda); L -2.1; DV-7.2, 12 degrees angle] (Rivera-Meza et al., 2014 (link)) according to the atlas of Paxinos and Watson (1998 ) (Paxinos and Watson, 1998 ). After recovery, animals were transferred to individual cages at the animal station, with access to water and food ad libitum. One week after surgery, the rats were infused each day with 100 ng of gp91 ds–tat (Anaspec) 20 min before exposure to CPP assay.
Apocynin (4′-Hydroxy-3′-methoxyacetophenone, Sigma) was added to drinking water at 5 mM and treatment was performed for 5 weeks (P30–P70) (Furukawa et al., 2004 (link); Harraz et al., 2008 (link); Espinosa et al., 2013 (link)).
Contreras M.L., de la Fuente-Ortega E., Vargas-Roberts S., Muñoz D.C., Goic C.A, & Haeger P.A. (2017). NADPH Oxidase Isoform 2 (NOX2) Is Involved in Drug Addiction Vulnerability in Progeny Developmentally Exposed to Ethanol. Frontiers in Neuroscience, 11, 338.
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8

Vascular Reactivity Assay Protocol

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ET-1 was from American Peptide (Sunnyvale, CA, USA) and Ang II was from MP Biomedicals (Solon, OH, USA). The thromboxane-prostanoid (TP) receptor antagonist[lS-[la,2a(Z),3a,4a]]-7-[3-[[2-[(phenylamino)carbonyl]hydrazino] methyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid (SQ 29,548) [23 (link)] and the NO synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME) were from Cayman Chemical (Ann Arbor, MI, USA), and the NADPH oxidase-selective inhibitor gp91ds-tat [24 (link)] was from Anaspec (Fremont, CA, USA). All other drugs were from Sigma-Aldrich (St. Louis, MO, USA). Stock solutions were prepared according to the manufacturer’s instructions, and diluted in physiological saline solution (PSS, composition in mmol/L: 129.8 NaCl, 5.4 KC1, 0.83 MgS04, 0.43 NaH2P04, 19 NaHC03, 1.8 CaCl2, and 5.5 glucose; pH 7.4) to the required concentrations before use.
Meyer M.R., Fredette N.C., Barton M, & Prossnitz E.R. (2014). Endothelin-1 but not Angiotensin II Contributes to Functional Aging in Murine Carotid Arteries. Life sciences, 118(2), 213-218.
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9

Investigating TRPV1 Knockout in Mice

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C57Bl/6 and TRPV1 knockout (KO) mice (C57Bl/6 background) were purchased from the Jackson Laboratory (Bar Harbor, ME). The ω-hydroxylase inhibitor N-hydroxy-N’-(4-n-butyl-2-methylphenyl)formamidine (HET0016) was purchased from Enzo (Farmingdale, NY). A784168 was purchased from Tocris (Minneapolis, MN). gp91 ds-tat and gp91 ds-tat scrambled were purchased from AnaSpec (Fremont, CA). The 20-HETE stable agonist N-(20-hydroxyeicosa-5[Z],14[Z]-dienoyl)glycine (20-5,14-HEDGE) was synthesized in the laboratory of J.R. Falck.
Zhang X., Demerdash N.E., Falck J.R., Munnuri S., Koehler R.C, & Yang Z.J. (2018). The contribution of TRPV1 channel to 20-HETE–aggravated ischemic neuronal injury. Prostaglandins & other lipid mediators, 137, 63-68.
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10

NOX2 Inhibitor gp91ds-tat Modulates Retinal Pressure

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The peptide gp91ds-tat (YGRKKRRQRRRCSTRIRRQL-NH2), a NOX2-specific inhibitor, interferes with NADPH oxidase assembly by targeting a sequence essential for binding gp91phox with p47phox [26 (link), 27 ]. HIV-tat peptide, an amino acid sequence internalized by all cells, is linked to the gp91phox sequence to facilitate cell entry [28 ]. For the Pressure 60 mmHg model, the retinal explants from C57BL/6J mice were incubated by adding different concentrations of gp91ds-tat (Anaspec, San Jose, CA, USA) in the standard culture medium for 24 h (Fig. 1C). Control or Pressure 60 mmHg retinas were incubated in the standard culture medium (Vehicle). For the in vivo H-IOP model, the different concentrations of gp91ds-tat (dissolved in normal saline) were injected intraperitoneally (10 ml/kg) 30 min after the H-IOP surgery and once every two days until the end of the experiment. Sham and H-IOP mice were injected with normal saline (Vehicle) (10 ml/kg). The mice were euthanized by cervical dislocation after two weeks. The preparation of retinal explants was performed as described above.
Shi X., Li P., Herb M., Liu H., Wang M., Wang X., Feng Y., van Beers T., Xia N., Li H, & Prokosch V. (2024). Pathological high intraocular pressure induces glial cell reactive proliferation contributing to neuroinflammation of the blood-retinal barrier via the NOX2/ET-1 axis-controlled ERK1/2 pathway. Journal of Neuroinflammation, 21, 105.
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