Thalamus tissue powder was sonicated in 1M Tris (pH 7.5) membrane lysis buffer containing 1M NaCl, 1% Trition X-100, 5mM EDTA, proteinase inhibitor and phosphatase inhibitor. Supernatant was collected after 10 min centrifugation at 18,000 × g, 4°C. Protein concentration was measured by Bradford assay. The western blot procedure was the same as previously published (Pang et al. 2013 (link)). Briefly, proteins were separated on 4–20% criterion TGX gels (BioRad, 567–1094) and transferred overnight to Immobilon PVDF transfer membranes (Millipore, Billerica, MA, IPFL00010). Membranes were blocked over night at 4°C with primary antibodies and blocked 1 hr at room temperature with secondary antibodies. Primary antibodies purchased from Abcam (Cambridge, MA) included rabbit anti-GFAP (ab7260, 1:50,000), rabbit anti-GGT7 (ab80903, 1:1000), mouse anti-MAP2 (ab11267, 1:4000), mouse anti-MRP1 (ab32574, 1:500), mouse anti-CD68 (ab31630, 1:500). Mouse anti-NeuN was from Millipore (Billerica, MA, MAB377, 1:1000) and mouse anti-β-actin was from Santa Cruz (Santa Cruz, CA, sc-8432, 1:1000). Rabbit anti-GPx4 antibody was purchased from Cayman (Ann Arbor, MI, 10005258, 1:300) and goat anti-GPx1 was from R&D system (Minneapolis, MN, AF3798, 1:300). Secondary antibodies were from Li-Cor (Lincoln, NE).
Goat anti gpx1
Manufactured by R&D Systems
Sourced in United States
Goat anti-GPx1 is an antibody product used in laboratory research. It is designed to detect and bind to the glutathione peroxidase 1 (GPx1) protein. GPx1 is an enzyme involved in the reduction of hydrogen peroxide and other organic hydroperoxides.
Automatically generated - may contain errors
Lab products found in correlation
Immobilon pvdf transfer membrane, by Merck Group (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Tgx gel, by Bio-Rad (1 mentions)
Ab11267, by Abcam (1 mentions)
Ab32574, by Abcam (1 mentions)
Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions)
Ab7260, by Abcam (1 mentions)
Abts substrate, by Thermo Fisher Scientific (1 mentions)
2 protocols using goat anti gpx1
1
Western Blot Analysis of Thalamus Proteins
2
Quantifying Antioxidant Enzyme Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Clarified supernatants (15 µg protein/mL; 100 µL) were prepared in coating buffer (containing 15 mM NaCO3 and 35 mM NaHCO3, pH 7.4), loaded onto a Maxisorp plate (Nalge Nunc, Rochester, USA) and incubated overnight at 4 °C. The plate was then blocked for 2 h in 1% w/v skim milk and incubated with primary antibody (3 h, 23 °C). Primary antibodies included goat anti-SOD1 (Santa Cruz, USA, #sc8637; dilution 1:200 v/v), goat anti-SOD2 (Sigma, USA, #S2147; 1:200 v/v) and goat anti-GPX1 (R&D Systems, USA, #AF3798; 1:500 v/v). Wells were then incubated with secondary antibody (1:5000 v/v dilution) (anti-mouse IgG-HRP and anti-rabbit IgG-HRP; 1 h at 20 °C). Next, a solution of ABTS substrate (Life Technologies, Carlsbad, CA) was added to each well followed by (1% w/v SDS) and monitoring at 410 nm using the FLUOstar Omega reader with values normalized to total protein.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!