The contralesional cortices washed with PBS was lysed with RIPA buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% NP40, 0.25% Na-deoxycholate, and 0.1% SDS) containing a protease inhibitor mixture and phosphatase inhibitors (Sigma-Aldrich, St. Louis, MO, USA). (BDNF, tPA, furin: Santa Cruz (R)) was added to SDS-PAGE and electroporated into Polyvinylidene fluoride membrane. Thirty micrograms of soluble protein were subjected to SDS–PAGE and electrotransferred onto a PVDF membrane. Specific protein bands were detected using specific antibodies (BDNF, tPA, furin: Santa Cruz biotechnology, Inc., Dallas, TX, USA, and β-actin: Sigma-Aldrich, St. Louis, MO, USA) and enhanced chemiluminescence (Pierce, Rockford, IL, USA) [55 (link)].
Furin
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Furin is a protease enzyme that cleaves protein precursors at specific sites. It is involved in the processing and activation of a variety of proteins.
Automatically generated - may contain errors
Lab products found in correlation
Skov3, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
P stat3, by Cell Signaling Technology (2 mentions)
P tyk2 tyr1054 1055, by Cell Signaling Technology (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Dmem media, by Thermo Fisher Scientific (2 mentions)
Tgf β, by Cell Signaling Technology (2 mentions)
P jak1 tyr1034 1035, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Mesothelin, by Cell Signaling Technology (2 mentions)
P erbb2, by Cell Signaling Technology (2 mentions)
Igf 1rβ, by Cell Signaling Technology (2 mentions)
Jak2 3230s, by Cell Signaling Technology (2 mentions)
P jak2 tyr1008, by Cell Signaling Technology (2 mentions)
Recombinant human neuregulin β 1 nrg1, by Thermo Fisher Scientific (2 mentions)
P erbb3, by Cell Signaling Technology (2 mentions)
β actin, by Merck Group (1 mentions)
Brain derived neurotrophic factor (bdnf), by Santa Cruz Biotechnology (1 mentions)
Cd147, by R&D Systems (1 mentions)
4 protocols using furin
1
Western Blot Analysis of BDNF, tPA, Furin
2
Histological Analysis of COVID-19 Receptor Proteins
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Formalin‐fixed and paraffin‐embedded lung tissue samples were cut into 2.5‐μm thick serial sections for histological analysis. Sections were deparaffinized in ROTICLEAR® (Carl Roth, Arlesheim, Switzerland), followed by decreasing concentrations of ethanol. Antigen retrieval was performed using citrate buffer (pH 9) in a 95 °C hot water bath for 20 min. Following blocking with 3% BSA in PBS for 1 h, sections were incubated with antibodies to ACE2 (1:100, #HPA000288; Sigma Aldrich, St Louis, MO, USA), TMPRSS2 (1:800, #bs‐6285R; Biossusa, Woburn, MA, USA), Ezrin (1:100, #3145S; Cell Signaling Technology, Boston, MA, USA), Furin (1:100, #sc‐133 142; Santa Cruz, Dallas, TX, USA), or CD147 (1:100, #AF972; R&D Systems, Minneapolis, MN, USA) at 4 °C overnight. The primary antibodies were detected with the immPRESS anti‐rabbit, anti‐goat, or anti‐mouse polymer detection kit using NovaRed peroxidase (HRP) substrate (Vector Laboratories, Burlingame, CA, USA). Hemalaun was used for nuclear counterstaining. Images were obtained using an Olympus VS120 slide scanning microscope (Olympus, Tokyo, Japan) at ×40 magnification.
3
Characterization of Ovarian Cancer Cell Lines
4
Characterization of Ovarian Cancer Cell Lines
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Ovarian cancer cell line OVCAR5 cells were purchased from the National Cancer Institute’s cell line repository. SKOV3, cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and HeyA8 cells were received from the characterized cell line core at MD Anderson Cancer Center. Cells were cultured in DMEM media (GIBCO, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS, Invitrogen Life Technologies, Carlsbad, CA) and 1% penicillin/streptomycin (GIBCO). ID8 cells (a kind gift from Dr. Weiguo Cui) were cultured in Dulbecco’s modified Eagle’s medium (GIBCO) supplemented with 10% FBS, 1% penicillin/streptomycin, 1% insulin (5 g/mL), 5 g/mL transferrin, 5 ng/mL sodium selenite (1X ITS; Sigma Cat. No. I3146). We obtained, ERBB2 (#2165), p-ERBB2 (Tyr1248)(#2247), ERBB3 (#12708), p-ERBB3 (Tyr1289), IGF1R-β (#3027), GAPDH (#5174), mesothelin (#99966S), TGF-β (#3711), STAT3 (#9139), p-STAT3 (Tyr705)(#9145), p-STAT3 (Ser727)(#34911), JAK1 (#3344S), p-JAK1(Tyr1034/1035) (#74129S), JAK2 (3230S), p-JAK2 (Tyr1008) (#8082S) TYK2 (#14193S) and p-TYK2 (Tyr1054/1055) (#68790S) from Cell Signaling Technology (Danvers, MA), Furin (#sc-133142) was obtained from Santa Cruz Biotechnology. Recombinant Human Neuregulin β−1 (NRG1), was purchased from PeproTech, Cat# 100–03.
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