RIPK1 antibody | knockout validation | BD Biosciences 610459

This is a knockout-validated antibody summary, based on the publications, "Biliary tract instillation of a SMAC mimetic induces TRAIL-dependent acute sclerosing cholangitis-like injury in mice" (Western blot, figure 4b) [2], "Combination of IAP antagonist and IFNγ activates novel caspase-10- and RIPK1-dependent cell death pathways" (Western blot, figure 4d) [3], "Kinase-independent functions of RIPK1 regulate hepatocyte survival and liver carcinogenesis" (Western blot, figure 1h) [4], "Caspase-8 scaffolding function and MLKL regulate NLRP3 inflammasome activation downstream of TLR3" [1] (as discussed below), "RIPK1 inhibits ZBP1-driven necroptosis during development" [5] (see supplementary figure 4a in the article), and "PELI1 functions as a dual modulator of necroptosis and apoptosis by regulating ubiquitination of RIPK1 and mRNA levels of c-FLIP" (Western blot, figure 3c) [6]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

RIPK1 antibody | knockout validation | BD Biosciences 610459 figure 1
Figure 1. Immunoblot of stable RIPK1 in macrophages derived from WT or RIPK1-KO mice. Caspase-1 is used as a control. From [1].
Antibody information

Antibody: RIPK1

Catalog number: 610459

Summary: Mouse IgG2a raised against human RIP aa. 385-650. Detects human, mouse, rat, dog, and chicken by western blot, immunofluorescence, immunoprecipitation. It is not recommended for immunohistochemistry.

Company: BD Biosciences

Validation Method

Western blot

Sample

macrophages derived from WT or RIPK1-KO mice

Primary incubation

1/1,000–1/2,000 dilution

Notes

Caspase-1 might not be a good housekeeping gene to use as a loading control, but authors conclude that it showed no changes.

References
  1. Kang S, Fernandes Alnemri T, Rogers C, Mayes L, Wang Y, Dillon C, et al. Caspase-8 scaffolding function and MLKL regulate NLRP3 inflammasome activation downstream of TLR3. Nat Commun. 2015;6:7515 pubmed publisher
  2. Guicciardi M, Krishnan A, Bronk S, Hirsova P, Griffith T, Gores G. Biliary tract instillation of a SMAC mimetic induces TRAIL-dependent acute sclerosing cholangitis-like injury in mice. Cell Death Dis. 2017;8:e2535 pubmed publisher
  3. Tanzer M, Khan N, Rickard J, Etemadi N, Lalaoui N, Spall S, et al. Combination of IAP antagonist and IFNγ activates novel caspase-10- and RIPK1-dependent cell death pathways. Cell Death Differ. 2017;24:481-491 pubmed publisher
  4. Van T, Polykratis A, Straub B, Kondylis V, Papadopoulou N, Pasparakis M. Kinase-independent functions of RIPK1 regulate hepatocyte survival and liver carcinogenesis. J Clin Invest. 2017;127:2662-2677 pubmed publisher
  5. Newton K, Wickliffe K, Maltzman A, Dugger D, Strasser A, Pham V, et al. RIPK1 inhibits ZBP1-driven necroptosis during development. Nature. 2016;540:129-133 pubmed publisher
  6. Wang H, Meng H, Li X, Zhu K, Dong K, Mookhtiar A, et al. PELI1 functions as a dual modulator of necroptosis and apoptosis by regulating ubiquitination of RIPK1 and mRNA levels of c-FLIP. Proc Natl Acad Sci U S A. 2017;114:11944-11949 pubmed publisher