Phospho-RIPK1 (Ser161) Antibody [A1A2]

Application: Reactivity: Human

Usage Information

Dilution
WB1:2000- 1:20000 IHC1:250-1:1000 IF1:400-1:1600 FCM1:23000

Application
WB, IHC, IF, FCM, ELISA

Reactivity
Human

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
76 kDa 74 kDa, 45 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Human small intestine tissue; Jurkat cells; Raji cells; THP-1 cells; U2OS cells; HEK-293T cells (Calyculin A treated); HeLa cells (Calyculin A treated)
Negative Control

Datasheet & SDS

Biological Description

Specificity
Phospho-RIPK1 (Ser161) Antibody [A1A2] detects endogenous levels of total RIPK1 protein only when it is phosphorylated at Ser161.

Clone
A1A2

Synonym(s)
Receptor-interacting serine/threonine-protein kinase 1; Cell death protein RIP; Receptor-interacting protein 1; RIP-1; RIPK1; RIP; RIP1

Background
Phospho-RIPK1 (Ser161) is the activated form of receptor-interacting serine/threonine-protein kinase 1 (RIPK1), a pseudokinase of the RIPK family, and marks autophosphorylation within the kinase activation loop, enabling conformational changes that license necroptosis and apoptosis downstream of TNFR1 and TLR3 signaling. Ser161 is located in the activation loop alongside Ser166 and Thr169; phosphorylation at S161 (which is hydrogen-bonded to Asp156 in the inactive state) stabilizes the open conformation of RIPK1, exposing the catalytic cleft for substrate access. Mutation S161N disrupts this H-bonding and reduces kinase activity, while the phosphomimetic S161E variant sustains necroptotic signaling. S161 autophosphorylation is a kinase activation checkpoint: it works cooperatively and redundantly with S166 phosphorylation to form the RIPK1 scaffold-to-death complex-II (RIPK1-FADD-caspase-8-RIPK3/MLKL) in the context of TAK1 or IKK inhibition. S161N mutation partially blocks keratinocyte necroptosis and skin inflammation in Ikk2^E-KO mice, while combined S161N/S166A mutations abolish both apoptosis and necroptosis, revealing functional redundancy; S161E can override S166A’s block on necroptosis. phospho-S161 transduces TNF-driven lethality (Ripk1^{S161N} mice are protected against TNFR1-induced shock), licenses RIPK3-MLKL pore formation, and sustains inflammation via NF-κB priming. Hyperactive S161 signaling drives inflammatory skin disorders such as psoriasis and dermatitis through keratinocyte death, while S161N knock-in prevents inflammation without causing immunosuppression.

Background

Phospho-RIPK1 (Ser161) is the activated form of receptor-interacting serine/threonine-protein kinase 1 (RIPK1), a pseudokinase of the RIPK family, and marks autophosphorylation within the kinase activation loop, enabling conformational changes that license necroptosis and apoptosis downstream of TNFR1 and TLR3 signaling. Ser161 is located in the activation loop alongside Ser166 and Thr169; phosphorylation at S161 (which is hydrogen-bonded to Asp156 in the inactive state) stabilizes the open conformation of RIPK1, exposing the catalytic cleft for substrate access. Mutation S161N disrupts this H-bonding and reduces kinase activity, while the phosphomimetic S161E variant sustains necroptotic signaling. S161 autophosphorylation is a kinase activation checkpoint: it works cooperatively and redundantly with S166 phosphorylation to form the RIPK1 scaffold-to-death complex-II (RIPK1-FADD-caspase-8-RIPK3/MLKL) in the context of TAK1 or IKK inhibition. S161N mutation partially blocks keratinocyte necroptosis and skin inflammation in Ikk2^E-KO mice, while combined S161N/S166A mutations abolish both apoptosis and necroptosis, revealing functional redundancy; S161E can override S166A’s block on necroptosis. phospho-S161 transduces TNF-driven lethality (Ripk1^{S161N} mice are protected against TNFR1-induced shock), licenses RIPK3-MLKL pore formation, and sustains inflammation via NF-κB priming. Hyperactive S161 signaling drives inflammatory skin disorders such as psoriasis and dermatitis through keratinocyte death, while S161N knock-in prevents inflammation without causing immunosuppression.

References
https://pubmed.ncbi.nlm.nih.gov/40996439/ https://pubmed.ncbi.nlm.nih.gov/40996440/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%