RIPA (Sigma, USA) reagents were used to extract protein from NPCs. Protein concentration was determined by a BCA kit (Sigma). Additionally, proteins (40 µg/per lane) were isolated by 15% SDS-PAGE gel. After that, the proteins were transferred onto PVDF membranes (Bio-rad, USA). The membranes were blocked with 5% skimmed milk for 2 h and incubated with primary antibodies including SLC7A11 (1:600, Abcam, USA, ab275411), GPX4 (1: 800, Abcam, USA, ab262509), ACSL4 (1: 1000, Abcam, USA, ab155282), and GAPDH (1: 1500, Abcam, USA, ab8245) at 4 °C overnight, followed by incubation with secondary goat anti-mouse antibody to immunoglobulin G (IgG; 1: 2000, Abcam, USA, ab150113) and goat anti-rabbit antibody to IgG (1:2000, Abcam, USA, ab150077) for 2 h. Finally, the protein expression was determined with an ECL kit. GAPDH was used as internal reference.
Ab275411
Manufactured by Abcam
Sourced in United States
Ab275411 is a laboratory product offered by Abcam. It is a tool used for research purposes. No further details about its core function or intended use can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab262509, by Abcam (2 mentions)
Ab8245, by Abcam (2 mentions)
Ab155282, by Abcam (1 mentions)
Ab150113, by Abcam (1 mentions)
Bca kit, by Merck Group (1 mentions)
Radioimmunoprecipitation assay (ripa), by Merck Group (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Ab150077, by Abcam (1 mentions)
Ab254360, by Abcam (1 mentions)
Ab215203, by Abcam (1 mentions)
Ab25901, by Abcam (1 mentions)
Ab205718, by Beyotime (1 mentions)
Ripa reagent, by Beyotime (1 mentions)
Ecl reagent, by Beyotime (1 mentions)
Bca kit, by Beyotime (1 mentions)
Ab263899, by Abcam (1 mentions)
2 protocols using ab275411
1
Western Blot Analysis of Ferroptosis Markers
2
Protein Expression Analysis in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were extracted using RIPA reagent (Beyotime), and their concentrations were determined using BCA kit (Beyotime), proteins were loaded to 10% SDS-PAGE for band separation. Separated bands were transferred to PVDF membranes and then blocked with 5% skim milk at 37 °C for 1 h. After that, these membranes containing protein bands were exposed to the primary antibodies overnight at 4 °C environment, such as anti-GPX4 (ab262509, 1:1000; Abcam, USA), anti-SLC7A11 (ab275411, 1:1000), anti-NLRP3 (ab263899, 1:1000), anti-cleaved caspase-1 (ab25901, 1:1000), anti-IL-1β (ab254360, 1:1000), anti-GSDMD-N (ab215203, 1:1000) and anti-GAPDH (ab8245, 1:1000). Next, these membranes were exposed to the corresponding secondary antibody conjugated with horseradich peroxidase (ab205718, 1:2000) at 37 °C for 2 h. Finally, the protein signaling was checked using the enhanced chemiluminescence (ECL) reagent (Beyotime, China), and the intensity of the protein bands was quantified using Image J software 10.0 (Bethesda, USA).
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!