Total ventricular extracts were prepared from the LV myocardium as described previously [31 (link)]. Protein concentrations were measured by BCA (Sigma-Aldrich, USA) assay. Equal amount of protein extracts was separated with SDS-page and transferred to a nitrocellulose membrane (Millipore, USA). The following primary antibodies were used in the present study: anti-GAPDH (USBiological, USA), anti-FZD1 (R&D systems, USA), anti-active-β-catenin (Millipore, USA), anti-β-catenin (Santa Cruz Biotechnology, USA), anti-p-GSK-3β (Cell-Signaling Technology, USA), anti-GSK-3β (BD Biosciences, Germany). HRP-conjugated anti-rabbit IgG, anti-mouse IgG, and anti-goat IgG antibodies (Santa Cruz Biotechnology, USA) were used as secondary antibodies. The blots were analyzed and quantified by densitometry using Image J program.
Anti fzd1
Manufactured by R&D Systems
Sourced in United States
Anti-Fzd1 is a recombinant protein that binds to the Frizzled-1 (Fzd1) receptor, a member of the Frizzled family of G protein-coupled receptors involved in the Wnt signaling pathway. The product is intended for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti β catenin, by Cell Signaling Technology (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Hrp conjugated anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Anti β catenin, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh, by US Biological (1 mentions)
Bicinchoninic acid (bca), by Merck Group (1 mentions)
Anti p gsk 3β, by Cell Signaling Technology (1 mentions)
Anti gsk 3β, by BD (1 mentions)
Anti active β catenin, by Merck Group (1 mentions)
Protease inhibitor and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Anti nlrp3, by Cell Signaling Technology (1 mentions)
Amersham protran 0.2 nc, by GE Healthcare (1 mentions)
Anti bdnf, by Abcam (1 mentions)
Anti gdnf, by Santa Cruz Biotechnology (1 mentions)
Anti p nf κb p65, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
Ecl kit, by Merck Group (1 mentions)
3 protocols using anti fzd1
1
Western Blot Analysis of Frizzled-1 Signaling
2
Quantifying Astrocyte Protein Signaling
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Protein from astrocytes was extracted on ice with RIPA buffer plus protease inhibitor and phosphatase inhibitor cocktail (both Thermo Fisher Scientific Inc.). Protein concentration was determined by BCA Protein Assay (Thermo Fisher Scientific Inc.). Protein extracts (30 µg) were separated by SDS-PAGE and transferred onto nitrocellulose membranes (AmershamProtran 0.2 NC; GE Healthcare Life Sciences, Chicago, IL, USA). The membranes were then incubated with anti-p-NF-κB/p65 (Cell Signaling Technology, Inc., Danvers, MA, USA), anti-Nrf2, anti-heme oxygenase-1 (HO-1; both Abcam, Cambridge, MA, USA), anti-Nlrp3, anti-PI3K, anti-p-Akt (all Cell Signaling Technology, Inc.), anti-WNT-1 (Abgent Inc., San Diego, CA, USA), anti-Fzd1 (R&D Systems, Inc., Minneapolis, MN, USA), anti-β-catenin (Cell Signaling Technology, Inc.), anti-BDNF (Abcam), anti-GDNF (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), and anti-β-actin (Cell Signaling Technology, Inc.) antibodies overnight at 4°C. Bands were visualized by HRP-conjugated secondary antibodies and chemiluminescence (ECL) kit (EMD Millipore, Billerica, MA, USA) under a ChemiDoc XRS+ system (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
3
Quantifying Neurobiological Markers in Mouse Brain
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The mouse brain was extracted using protein extraction buffer (T-PER, Thermo Fisher, USA) containing protease and phosphatase inhibitors (Thermo Fisher, USA). Equal amounts of protein extracts (20µg) were separated by SDS-PAGE and transferred onto a nitrocellulose lter (NC) membrane (GE Healthcare Life Sciences, USA). After being blocked with 5% non-fat milk, membranes were incubated at 4°C overnight with primary antibodies (all at 1:1000) as follows: anti-β-actin (Abcam, USA), anti-p-mTOR(Abcam, USA), anti-GDNF(Abcam, USA), anti-BNDF(Abcam, USA), anti-CNTF(Abcam, USA), anti-Bcl-2(Abcam, USA), anti-Bax (Abcam, USA), anti-p110α-PI3K (C73F8) (Cell Signaling Technology, MA), anti-p-Akt (Ser473) (Cell Signaling Technology, MA), anti-β-catenin (Cell Signaling Technology, MA), anti-Fzd1 (R&D system, USA), anti-NLRP3 (R&D system, USA), anti-WNT1 (ABGENT, USA), anti-iNOS/NOS(BD, USA) or anti-Arginase1 (BD, USA). After being washed in TBST, the immunoblots underwent incubation with horseradish peroxidase-conjugated secondary antibodies (Jackson Lab, USA) for 1h. Bands were visualized and analyzed using a ChemiDoc XRS (Bio-Rad, USA) and Image Lab(Bio-Rad, USA).
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