RIP3 antibody | knockout validation | Cell Signaling 95702

This is a knockout-validated antibody summary, based on publications "Melatonin attenuates chronic pain related myocardial ischemic susceptibility through inhibiting RIP3-MLKL/CaMKII dependent necroptosis", as cited below [1], and "Microbial signals, MyD88, and lymphotoxin drive TNF-independent intestinal epithelial tissue damage" for western blot knockout validation (figure 6d) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: RIP3

Catalog number: 95702

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Val370 of mouse RIP3 protein. Reacts with mouse by western blot, immunoprecipitation, immunofluorescence (immunocytochemistry) and flow cytometry.

Validation Method

Western blot

Sample

Heart tissue from wild-type and RIP3 KO mice.

Blocking agent

3% nonfat milk in PBST for 1 h.

Primary incubation

At 4 °C over the night.

Secondary incubation

1:5,000 dilution horseradish peroxidase-conjugated antibodies for 1 h at room temperature.

Detection

Chemiluminescence reagents (Millipore, Billerica, MA, USA) was used to visualize the blots.

Figure

Representative western blots for myocardial RIP3 expression in RIP3 knockout (RIP3 KO) mice and their wild-type (WT) littermates. Please see Figure 3b in the article [1].

References
  1. Yang Z, Li C, Wang Y, Yang J, Yin Y, Liu M, et al. Melatonin attenuates chronic pain related myocardial ischemic susceptibility through inhibiting RIP3-MLKL/CaMKII dependent necroptosis. J Mol Cell Cardiol. 2018;125:185-194 pubmed publisher
  2. Rusu I, Mennillo E, Bain J, Li Z, Sun X, Ly K, et al. Microbial signals, MyD88, and lymphotoxin drive TNF-independent intestinal epithelial tissue damage. J Clin Invest. 2022;132: pubmed publisher