Product: Phospho-RIPK2 (Ser176) Antibody
Catalog: AF0049
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Mol.Wt.: 70kD; 61kD(Calculated).
Uniprot: O43353
RRID: AB_2834094

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
Phospho-RIPK2 (Ser176) Antibody detects endogenous levels of RIPK2 only when phosphorylated at Sersine 176.
RRID:
AB_2834094
Cite Format: Affinity Biosciences Cat# AF0049, RRID:AB_2834094.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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

CARD 3; CARD carrying kinase; CARD containing ICE associated kinase; CARD containing IL 1 beta ICE kinase; CARD containing IL1 beta ICE kinase; CARD containing interleukin 1 beta converting enzyme (ICE) associated kinase; CARD containing interleukin 1 beta converting enzyme associated kinase; CARD-containing IL-1 beta ICE-kinase; CARD-containing interleukin-1 beta-converting enzyme-associated kinase; CARD3; CARDIAK; CCK; CLARP kinase; GIG 30; GIG30; Growth inhibiting gene 30; Receptor interacting protein (RIP) like interacting caspase like apoptosis regulatory protein (CLARP) kinase; Receptor interacting protein 2; Receptor interacting serine threonine kinase 2; Receptor interacting serine/threonine protein kinase 2; Receptor-interacting protein 2; Receptor-interacting serine/threonine-protein kinase 2; RICK; RIP 2; RIP like interacting CLARP kinase; RIP-2; RIP-like-interacting CLARP kinase; RIPK 2; Ripk2; RIPK2_HUMAN; TNFRSF; Tyrosine-protein kinase RIPK2; UNQ277/PRO314/PRO34092;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O43353 RIPK2_HUMAN:

Detected in heart, brain, placenta, lung, peripheral blood leukocytes, spleen, kidney, testis, prostate, pancreas and lymph node.

Description:
RIPK2 a tyrosine kinase-like kinase of the RIPK family. Activates pro-caspase-1 and pro-caspase-8. Potentiates casp-8-mediated apoptosis. May activate NF-kappaB. Note: This description may include information from UniProtKB.
Sequence:
MNGEAICSALPTIPYHKLADLRYLSRGASGTVSSARHADWRVQVAVKHLHIHTPLLDSERKDVLREAEILHKARFSYILPILGICNEPEFLGIVTEYMPNGSLNELLHRKTEYPDVAWPLRFRILHEIALGVNYLHNMTPPLLHHDLKTQNILLDNEFHVKIADFGLSKWRMMSLSQSRSSKSAPEGGTIIYMPPENYEPGQKSRASIKHDIYSYAVITWEVLSRKQPFEDVTNPLQIMYSVSQGHRPVINEESLPYDIPHRARMISLIESGWAQNPDERPSFLKCLIELEPVLRTFEEITFLEAVIQLKKTKLQSVSSAIHLCDKKKMELSLNIPVNHGPQEESCGSSQLHENSGSPETSRSLPAPQDNDFLSRKAQDCYFMKLHHCPGNHSWDSTISGSQRAAFCDHKTTPCSSAIINPLSTAGNSERLQPGIAQQWIQSKREDIVNQMTEACLNQSLDALLSRDLIMKEDYELVSTKPTRTSKVRQLLDTTDIQGEEFAKVIVQKLKDNKQMGLQPYPEILVVSRSPSLNLLQNKSM

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

PTMs - O43353 As Substrate

Site PTM Type Enzyme
K17 Ubiquitination
Y23 Phosphorylation
R26 Methylation
S168 Phosphorylation
S174 Phosphorylation
S176 Phosphorylation O43353 (RIPK2)
S178 Phosphorylation
S180 Phosphorylation
S181 Phosphorylation
K182 Ubiquitination
K203 Ubiquitination
K209 Ubiquitination
S319 Phosphorylation
S345 Phosphorylation
S357 Phosphorylation
S363 Phosphorylation
S374 Phosphorylation
Y381 Phosphorylation P12931 (SRC)
K384 Methylation
S393 Phosphorylation
S399 Phosphorylation
K410 Ubiquitination
T411 Phosphorylation
T412 Phosphorylation
S428 Phosphorylation
Y474 Phosphorylation O43353 (RIPK2)
K480 Ubiquitination
K508 Ubiquitination
S527 Phosphorylation
S529 Phosphorylation
S531 Phosphorylation
K538 Ubiquitination
S539 Phosphorylation

PTMs - O43353 As Enzyme

Substrate Site Source
O43353 (RIPK2) S176 Uniprot
O43353 (RIPK2) Y474 Uniprot
Q13568 (IRF5) S435 Uniprot
Q13568 (IRF5) S446 Uniprot

Research Backgrounds

Function:

Serine/threonine/tyrosine kinase that plays an essential role in modulation of innate and adaptive immune responses. Upon stimulation by bacterial peptidoglycans, NOD1 and NOD2 are activated, oligomerize and recruit RIPK2 through CARD-CARD domains. Contributes to the tyrosine phosphorylation of the guanine exchange factor ARHGEF2 through Src tyrosine kinase leading to NF-kappaB activation by NOD2. Once recruited, RIPK2 autophosphorylates and undergoes 'Lys-63'-linked polyubiquitination by E3 ubiquitin ligases XIAP, BIRC2 and BIRC3. The polyubiquitinated protein mediates the recruitment of MAP3K7/TAK1 to IKBKG/NEMO and induces 'Lys-63'-linked polyubiquitination of IKBKG/NEMO and subsequent activation of IKBKB/IKKB. In turn, NF-kappa-B is released from NF-kappa-B inhibitors and translocates into the nucleus where it activates the transcription of hundreds of genes involved in immune response, growth control, or protection against apoptosis. Plays also a role during engagement of the T-cell receptor (TCR) in promoting BCL10 phosphorylation and subsequent NF-kappa-B activation. Plays a role in the inactivation of RHOA in response to NGFR signaling.

PTMs:

Autophosphorylated. Autophosphorylation at Tyr-474 is necessary for effective NOD2 signaling. Phosphorylated. Phosphorylation at Tyr-381 by Src kinase CSK occurs in a ARHGEF2-dependent manner and is required for NOD2-dependent innate immune activation.

Ubiquitinated on Lys-209; undergoes 'Lys-63'-linked polyubiquitination catalyzed by ITCH. Polyubiquitinated with 'Lys-48' and 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2, leading to activation of NF-kappa-B. Also undergoes 'Met-1'-linked polyubiquitination; the head-to-tail linear polyubiquitination is mediated by the LUBAC complex in response to NOD2 stimulation. Linear polyubiquitination is restricted by FAM105B/otulin, probably to limit NOD2-dependent proinflammatory signaling activation of NF-kappa-B.

Subcellular Location:

Cytoplasm.

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

Detected in heart, brain, placenta, lung, peripheral blood leukocytes, spleen, kidney, testis, prostate, pancreas and lymph node.

Subunit Structure:

Found in a signaling complex consisting of at least ARHGEF2, NOD2 and RIPK2. Interacts with ARHGEF2; the interaction mediates tyrosine phosphorylation of RIPK2 by Src kinase CSK. Binds to CFLAR/CLARP and CASP1 via their CARD domains. Binds to BIRC3/c-IAP1 and BIRC2/c-IAP2, TRAF1, TRAF2, TRAF5 and TRAF6. May be a component of both the TNFRSF1A and TNRFSF5/CD40 receptor complex. Interacts with NOD1. Interacts (via CARD domain) with NOD2 (via CARD domain). Interacts with MAP3K4; this interaction sequesters RIPK2 from the NOD2 signaling pathway. Interacts with IKBKG/NEMO. The polyubiquitinated protein interacts with MAP3K7/TAK1. Interacts with XIAP/BIRC4. Interacts with NLRP10. Interacts with CARD9. Interacts with INAVA; the interaction takes place upon PRR stimulation. Interacts (via CARD domain) with NGFR (via death domain).

Family&Domains:

Contains an N-terminal kinase domain and a C-terminal caspase activation and recruitment domain (CARD) that mediates the recruitment of CARD-containing proteins.

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

Research Fields

· Human Diseases > Infectious diseases: Bacterial > Shigellosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

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

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

References

1). Gao Q et al. Sodium valproate attenuates the iE-DAP induced inflammatory response by inhibiting the NOD1-NF-κB pathway and histone modifications in bovine mammary epithelial cells. Int Immunopharmacol 2020 Mar 14;83:106392 (PubMed: 32182568) [IF=3.943]

Application: WB    Species: bovine    Sample: BMECs

Fig. 4. |Protein expression of NOD1-mediated NF-κB signaling pathway genes and the expression and location of NOD1 and NF-κB p65 protein in BMECs. The cells were treated with iE‐DAP (10 μg/mL) or VPA (0.5 mmol/L) or pretreatment with VPA (0.5 mmol/L) followed by 10 μg/mL of iE‐DAP alone. Western blot analysis showing the NOD1, RIPK2 (A, B), p65, p-p65, IκB and p-IκB (C, D) levels. The protein expression was normalized by the respective abundance of β-actin and βtubulin.

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