Cautionary notes on the use of NF-κB p65 and p50 antibodies for CNS studies, Journal of Neuroinflammation
Por um escritor misterioso
Last updated 02 abril 2025

Background The characterization and cellular localization of transcription factors like NF-κB requires the use of antibodies for western blots and immunohistochemistry. However, if target protein levels are low and the antibodies not well characterized, false positive data can result. In studies of NF-κB activity in the CNS, antibodies detecting NF-κB proteins have been used to support the finding that NF-κB is constitutively active in neurons, and activity levels are further increased by neurotoxic treatments, glutamate stimulation, or elevated synaptic activity. The specificity of the antibodies used was analyzed in this study. Methods Selectivity and nonselectivity of commonly used commercial and non-commercial p50 and p65 antibodies were demonstrated in western blot assays conducted in tissues from mutant gene knockout mice lacking the target proteins. Results A few antibodies for p50 and p65 each mark a single band at the appropriate molecular weight in gels containing proteins from wildtype tissue, and this band is absent in proteins from knockout tissues. Several antibodies mark proteins that are present in knockout tissues, indicating that they are nonspecific. These include antibodies raised against the peptide sequence containing the nuclear localization signals of p65 (MAB3026; Chemicon) and p50 (sc-114; Santa Cruz). Some antibodies that recognize target proteins at the correct molecular weight still fail in western blot analysis because they also mark additional proteins and inconsistently so. We show that the criterion for validation by use of blocking peptides can still fail the test of specificity, as demonstrated for several antibodies raised against p65 phosphorylated at serine 276. Finally, even antibodies that show specificity in western blots produce nonspecific neuronal staining by immunohistochemistry. Conclusions We note that many of the findings in the literature about neuronal NF-κB are based on data garnered with antibodies that are not selective for the NF-κB subunit proteins p65 and p50. The data urge caution in interpreting studies of neuronal NF-κB activity in the brain.

The Role of NF-κB in Neuroinflammation

IκBβ is a positive and negative regulator of NF-κB activity during inflammation

Figure 1.5 from Is there a role for NF-κB in the suprachiasmatic circadian clock?

NFκB-Activated Astroglial Release of Complement C3 Compromises Neuronal Morphology and Function Associated with Alzheimer's Disease - ScienceDirect

Pathogens, Free Full-Text

NF-KappaB p50-p65 Pathway

Spinal NF-kB upregulation contributes to hyperalgesia in a rat model of advanced osteoarthritis - Yunze Li, Yixin Yang, Jinwan Guo, Xuejiao Guo, Zhiying Feng, Xuli Zhao, 2020

Frontiers Estrogen Receptor Beta Influences the Inflammatory p65 Cistrome in Colon Cancer Cells

Nuclear Factor-κB Contributes to Neuron-Dependent Induction of Glutamate Transporter-1 Expression in Astrocytes

β2-Adrenergic receptors in immunity and inflammation: Stressing NF-κB - ScienceDirect

Nuclear Factor κB Signaling in Atherogenesis Arteriosclerosis, Thrombosis, and Vascular Biology
Recomendado para você
-
fase 372 brain test02 abril 2025
-
Optical Illusion Brain Test: If you have Sharp Eyes Find the Number 372 in 20 Secs - News02 abril 2025
-
Super Danish Me – chasing hygge02 abril 2025
-
End-Stage Heart Disease and Indications for Heart Transplantation02 abril 2025
-
What is Adderall? — TACO Inc02 abril 2025
-
A Genetic Screen Implicates miRNA-372 and miRNA-373 As Oncogenes in Testicular Germ Cell Tumors: Cell02 abril 2025
-
A sense of self02 abril 2025
-
Arithmetic & Geometric Sequences, 154 plays02 abril 2025
-
A Genetic Screen Implicates miRNA-372 and miRNA-373 As Oncogenes in Testicular Germ Cell Tumors - ScienceDirect02 abril 2025
-
Think about your thoughts #stereo02 abril 2025
você pode gostar
-
Arizona football vs Oregon State: Players of the Game - Arizona02 abril 2025
-
Pokemoner - 🤜Download Pokemon Legends Arceus GBA - Team02 abril 2025
-
General Qui-Gon Jinn - if he had fought in the Clone Wars : r/StarWars02 abril 2025
-
resident evil mr x original|TikTok Search02 abril 2025
-
JOGO COMPLETO: SUÉCIA X ITÁLIA, 2ª RODADA02 abril 2025
-
Jogos de amanhã da Copa do Mundo 2022: horários do dia 04/12, Copa do Mundo02 abril 2025
-
Tema de videogame cenário fotografia desenho animado controlador de jogos grafite pixel meninos decoração de sala de jogos fundo festa sessão de fotos, 102 abril 2025
-
Luas Superiores VS. Hashiras (feat. Kaito Rapper, ÉoDan, Basara02 abril 2025
-
O Que Você Quer Ser Quando Crescer?02 abril 2025
-
MSI Nvidia GeForce RTX 4080 16GB GAMING X TRIO Graphics Card02 abril 2025