Product: RIPK1 Antibody
Catalog: AF7877
Description: Rabbit polyclonal antibody to RIPK1
Application: WB
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 76kDa; 76kD(Calculated).
Uniprot: Q13546
RRID: AB_2844241

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(86%), Bovine(86%), Horse(86%), Sheep(86%), Rabbit(86%), Dog(86%), Chicken(86%)
Clonality:
Polyclonal
Specificity:
RIPK1 Antibody detects endogenous levels of total RIPK1.
RRID:
AB_2844241
Cite Format: Affinity Biosciences Cat# AF7877, RRID:AB_2844241.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Cell death protein RIP; FLJ39204; OTTHUMP00000039163; Receptor (TNFRSF) interacting serine threonine kinase 1; receptor interacting protein 1; Receptor interacting protein; Receptor interacting protein kinase 1; Receptor interacting serine threonine protein kinase 1; Receptor TNFRSF interacting serine threonine kinase 1; Receptor-interacting protein 1; Receptor-interacting serine/threonine-protein kinase 1; Rinp; RIP 1; RIP; Rip-1; RIP1; RIPK 1; Ripk1; RIPK1_HUMAN; Serine threonine protein kinase RIP; Serine/threonine-protein kinase RIP;

Immunogens

Immunogen:

A synthesized peptide derived from human RIPK1, corresponding to a region within the internal amino acids.

Uniprot:
Gene(ID):
Sequence:
MQPDMSLNVIKMKSSDFLESAELDSGGFGKVSLCFHRTQGLMIMKTVYKGPNCIEHNEALLEEAKMMNRLRHSRVVKLLGVIIEEGKYSLVMEYMEKGNLMHVLKAEMSTPLSVKGRIILEIIEGMCYLHGKGVIHKDLKPENILVDNDFHIKIADLGLASFKMWSKLNNEEHNELREVDGTAKKNGGTLYYMAPEHLNDVNAKPTEKSDVYSFAVVLWAIFANKEPYENAICEQQLIMCIKSGNRPDVDDITEYCPREIISLMKLCWEANPEARPTFPGIEEKFRPFYLSQLEESVEEDVKSLKKEYSNENAVVKRMQSLQLDCVAVPSSRSNSATEQPGSLHSSQGLGMGPVEESWFAPSLEHPQEENEPSLQSKLQDEANYHLYGSRMDRQTKQQPRQNVAYNREEERRRRVSHDPFAQQRPYENFQNTEGKGTAYSSAASHGNAVHQPSGLTSQPQVLYQNNGLYSSHGFGTRPLDPGTAGPRVWYRPIPSHMPSLHNIPVPETNYLGNTPTMPFSSLPPTDESIKYTIYNSTGIQIGAYNYMEIGGTSSSLLDSTNTNFKEEPAAKYQAIFDNTTSLTDKHLDPIRENLGKHWKNCARKLGFTQSQIDEIDHDYERDGLKEKVYQMLQKWVMREGIKGATVGKLAQALHQCSRIDLLSSLIYVSQN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
86
Horse
86
Bovine
86
Sheep
86
Dog
86
Chicken
86
Rabbit
86
Xenopus
67
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

Serine-threonine kinase which is a key regulator of both cell death and cell survival. Exhibits kinase activity-dependent functions that trigger cell death and kinase-independent scaffold functions regulating inflammatory signaling and cell survival. Initiates ripoptocide which describes cell death that is dependent on RIPK1, be it apoptosis or necroptosis. Upon binding of TNF to TNFR1, RIPK1 is recruited to the TNF-R1 signaling complex (TNF-RSC also known as complex I) where it acts as a scaffold protein promoting cell survival, in part, by activating the canonical NF-kB pathway (By similarity). Specific conditions can however activate RIPK1, and its kinase activity then regulates assembly of two death-inducing complexes, namely complex IIa (RIPK1-FADD-CASP8) and the complex IIb (RIPK1-RIPK3-MLKL) and these complexes respectively drive apoptosis or necroptosis, a regulated form of necrosis. During embryonic development suppresses apoptosis and necroptosis and prevents the interaction of TRADD with FADD thereby limiting aberrant activation of CASP8 (By similarity). Phosphorylates DAB2IP at 'Ser-728' in a TNF- alpha-dependent manner, and thereby activates the MAP3K5-JNK apoptotic cascade. Required for ZBP1-induced NF-kappaB activation and activation of NF-kappaB by DNA damage and IR (By similarity).

PTMs:

Proteolytically cleaved by CASP8 at Asp-324. Cleavage is crucial for limiting apoptosis and necroptosis during embryonic development (By similarity). Cleavage abolishes NF-kappa-B activation and enhances the interaction of TRADD with FADD.

RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation. Phosphorylation of Ser-161 by RIPK3 is necessary for the formation of the necroptosis-inducing complex. Phosphorylation at Ser-25 represses its kinase activity and consequently prevents TNF-mediated RIPK1-dependent cell death. Phosphorylated at Ser-320 by MAP3K7 which requires prior ubiquitination with 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2 (By similarity). This phosphorylation positively regulates RIPK1 interaction with RIPK3 to promote necroptosis but negatively regulates RIPK1 kinase activity and its interaction with FADD to mediate apoptosis (By similarity).

Ubiquitinated with 'Lys-11'-, 'Lys-48'-, 'Lys-63'- and linear-linked type ubiquitin Ref.33). Polyubiquitination with 'Lys-63'-linked chains by TRAF2 induces association with the IKK complex. Deubiquitination of 'Lys-63'-linked chains and polyubiquitination with 'Lys-48'-linked chains by TNFAIP3 leads to RIPK1 proteasomal degradation and consequently down-regulates TNF-alpha-induced NFkappa-B signaling. 'Lys-48'-linked polyubiquitination by RFFL or RNF34 also promotes proteasomal degradation and negatively regulates TNF-alpha-induced NF-kappa-B signaling Ref.33). Linear polyubiquitinated; the head-to-tail linear polyubiquitination ('Met-1'-linked) is mediated by the LUBAC complex and decreases protein kinase activity. Deubiquitination of linear polyubiquitin by CYLD promotes the kinase activity (By similarity). Polyubiquitinated with 'Lys-48' and 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2, leading to activation of NF-kappa-B. Ubiquitinated with 'Lys-63'-linked chains by PELI1. Ubiquitination at Lys-377 with 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2 is essential for its phosphorylation at Ser-320 mediated by MAP3K7 (By similarity). This ubiquitination is required for NF-kB activation, suppresses RIPK1 kinase activity and plays a critical role in preventing cell death during embryonic development (By similarity).

Subcellular Location:

Cytoplasm. Cell membrane.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Family&Domains:

The death domain mediates dimerization and activation of its kinase activity during necroptosis and apoptosis (PubMed:29440439). It engages other DD-containing proteins as well as a central (intermediate) region important for NF-kB activation and RHIM-dependent signaling (PubMed:10356400).

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

References

1). Extracellular vesicles of Fusobacterium nucleatum compromise intestinal barrier through targeting RIPK1-mediated cell death pathway. Gut Microbes, 2021 (PubMed: 33769187) [IF=12.2]

Application: WB    Species: human    Sample: Caco2 cells

Figure 2. |FnEVs promote TNF-α induced Caco2 cells death and barrier loss. (a) Diagram of groups. (b) Apoptosis was analyzed by using the annexin V FITC/PI assay. (c) Bar graphs show the relative mean of Caco2 cells apoptosis rate in different view fields. (d) The releasing levels of LDH were detected. (e) Representative images of TUNEL stainings of Caco-2 cells (green, TUNEL positive; blue, DAPI). Scale bar = 50 um. (f) Immunoblot analysis of protein extracts from Caco2 cells with the indicated antibodies

Application: IHC    Species: mouse    Sample: colon

Figure 6. |RIPK1 mediated epithelial cell death drives exacerbated barrier loss in FnEVs-treated colitis mice. (a) Representative images of TUNEL stainings of colon sections on day 3 after colitis induction (red, TUNEL positive; blue, DAPI). Scale bar = 50 um (up) or 20 um (down). (b) Representative images of immunohistochemical stainings of FADD, RIPK1 and cCASP3 in the colon sections on day 3 after colitis induction. Scale bar = 50 um or 20 um (downmost).

2). Integrated Studies on Male Reproductive Toxicity of Decabromodiphenyl Ethane in Zebrafish Spermatozoa Ex Vivo, Male Zebrafish in Vivo, and GC-1 Cells in Vitro. Environmental health perspectives, 2024 (PubMed: 39570742) [IF=10.1]

3). Circular RNA FEACR inhibits ferroptosis and alleviates myocardial ischemia/reperfusion injury by interacting with NAMPT. Journal of biomedical science, 2023 (PubMed: 37370086) [IF=9.0]

4). Zinc oxide nanoparticles disrupt the mammary epithelial barrier via Z-DNA binding protein 1-triggered PANoptosis. Ecotoxicology and environmental safety, 2024 (PubMed: 39053182) [IF=6.2]

5). PTTG1 alleviates acute alcoholic liver injury by inhibiting endoplasmic reticulum stress‐induced hepatocyte pyroptosis. LIVER INTERNATIONAL, 2023 (PubMed: 36737842) [IF=6.0]

6). Tanshinone I exerts cardiovascular protective effects in vivo and in vitro through inhibiting necroptosis via Akt/Nrf2 signaling pathway. Chinese Medicine, 2021 (PubMed: 34183021) [IF=5.3]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 2 TI ameliorated t‑BHP induced cell necroptosis via RIP1/RIP3/MLKL pathway. a H9c2 cells were cultured with Nec-1 for 12 h. b Cells were exposed to t-BHP (150 μM) for 10 h after treated with Nec-1 for 2 h. c Cells were exposed to t-BHP (150 μM) for 10 h after treated with TI (1 μM) or Nec-1 (100 μM) for 2 h. MTT was employed to detect cell viability. d Cells were treated with t-BHP (150 μM) for 10 h when pretreated with TI (1 μM) or Nec-1 (100 μM) for 2 h, the LDH level was monitored by LDH kit. e–h Pretreated with TI (0.25, 0.5, and 1 μM) or Nec-1 (100 μM) for 2 h respectively, then exposed to t-BHP (150 μM) for 4 h, the protein expression was determined by western blotting. n = 3. *p < 0.05, **p < 0.01, ***p < 0.001 vs. t-BHP group

7). Sacubitril Ameliorates Cardiac Fibrosis Through Inhibiting TRPM7 Channel. Frontiers in Cell and Developmental Biology, 2021 (PubMed: 34778271) [IF=4.6]

8). Transient Inhibition of mTORC1 Signaling Ameliorates Irradiation-Induced Liver Damage. Frontiers in Physiology, 2019 (PubMed: 30984007) [IF=4.0]

Application: WB    Species: mouse    Sample: Liver

FIGURE 2 | Irradiation-induced cellular apoptosis in liver tissues. (A and B) Irradiated liver tissues were collected at days 1 and 3 after TBI. Representative pictures by TUNEL staining are shown (A, n = 6). Scale bar = 50 μm. Arrows indicate TUNEL-positive cells. The ratios of apoptotic cells in each field are presented in (B) (N.D: non-detected). (C and D) RIPK1 expression in liver tissues (n = 3). Expression of RIPK1 in liver tissues was detected by western blotting (C). GAPDH was used as a housekeeping control. Expression of RIPK1 was quantitated by ImageJ software (D).

9). ERAD deficiency promotes mitochondrial dysfunction and transcriptional rewiring in human hepatic cells. JOURNAL OF BIOLOGICAL CHEMISTRY, 2020 (PubMed: 32978261) [IF=4.0]

10). Sugarcane leaf polysaccharide exerts a therapeutic effect on cardiovascular diseases through necroptosis. Heliyon, 2023 (PubMed: 38027563) [IF=4.0]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 5 SLP alleviated TBHP-stimulated cell necroptosis through the RIP1/RIP3/MLKL signaling pathway in H9c2 cells. (A) H9c2 cells were incubated with SLP (25, 50, and 100 μg/mL) and then treated with TBHP (100 μM). The protein expression of p-RIP1, RIP1, p-RIP3, RIP3, p-MLKL, and MLKL was determined using western blotting. (B–D) Statistical analysis of the above protein expressions. n = 3. ##p < 0.01, ###p < 0.001, vs. the control group. *p < 0.05, **p < 0.01, ***p < 0.001, vs. TBHP group.

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