Rabbit RIP3 Polyclonal Antibody | anti-Ripk3 antibody

RIP3 Antibody

Cat. Num. AAA150424

• Gene Names: Ripk3; Rip3; AW107945; 2610528K09Rik
• Reactivity: Human, Mouse, Rat
• Applications: ELISA, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
• Purity: RIP3 Antibody is affinity chromatography purified via peptide column.
• Synonyms: RIP3; Polyclonal Antibody; RIP3 Antibody; Rip3; AW107945; 2610528K09Rik; RIP-like protein kinase 3; RIP-3; receptor-interacting serine-threonine kinase 3; anti-Ripk3 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Polyclonal
• Isotype: IgG
• Purity/Purification: RIP3 Antibody is affinity chromatography purified via peptide column.
• Form/Format: Liquid
• Concentration: 1 mg/ml (varies by lot)
• Sequence Length: 486

• Applicable Applications for anti-Ripk3 antibody: ELISA, IF, IHC-P, IP, WB
• Application Notes: WB: 0.1-0.5 ug/mL; IHC-P: 5 ug/mL; IF: 20 ug/mL; IP: 20 ug/mL; ; Antibody validated: Western Blot in mouse and human samples; Immunohistochemistryin mouse and rat samples; Immunofluorescence in mouse and rat samples;Immunoprecipitation in human samples; Immunogold EM in mouse samples. All otherapplications and species not yet tested
• Conjugate: Unconjugated
• Immunogen: Anti-RIP3 antibody (MBS150424) was raised against a peptide corresponding to 14 amino acids near the carboxy terminus of murine RIP3.The immunogen is located within the last 50 amino acids of RIP3.
• Buffer: RIP3 Antibody is supplied in PBS containing 0.02% sodium azide.
• Isoforms: Mouse RIP3 has one isoform (486aa, 53kD). Human RIP3 has 3 isoforms, including isoform1 (518aa, 57kD), isoform 2 (252aa, 28kD) and isoform 3 (231aa, 25kD). Rat RIP3 also hasone isoform (478aa, 52kD). MBS150424 can detect can detect isoforms of mouse and rat as well as human isoform 1.
• Validation: KO Validation; (Figure 7,9,13) shows RIP3 expression detected by anti-RIP3 antibodies (MBS150424) was disrupted in multiple tissues of RIP3 KO mice (figure 7, 13) and in RIP3 KO MEF cells (figure 9).; KD Validation; (Figure 10): Anti-RIP3 antibody (MBS150424) specificity was further verified by RIP3 specific knockdown. RIP3 signal in L929 cells transfected with RIP3 shRNA was disrupted in comparison with that in cells transfected with control shRNA.; Regulated Expression Validation; (Figure 7,11,13): RIP3 expression detected by anit-RIP3 antibodies (MBS150424) was up-regulated by treatment of cerulein (figure 7), LPS (figure 11) or acetaminophen (figure 13) in different tissues or cells.; Overexpression Validation; (Figure 12): RIP3 overexpression detected by anit-RIP3 antibodies (MBS150424) was observed in three cell lines stably transfected with RIP3.
• Positive Control: 1) Cat. No. MBS151754 – A431 Cell Lysate; 2) Cat. No. MBS151593 - 3T3/NIH Cell Lysate; 3) Cat. No. MBS151658- C2C12 Cell Lysate; 4) Cat. No. MBS151757- 3T3/Balb Cell Lysate
• Preparation and Storage: RIP3 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Western Blot (WB)

(Loading: 15 ?g of lysates per lane. Antibodies: RIP3 (MBS150424), (0.5 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution)

Western Blot (WB)

(Loading: 15 ug of lysates per lane. Antibodies: RIP3 MBS150424, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: MBS150424, 0.1 ug/mL in the presence of peptide blocking Lane 2: MBS150424, 0.1 ug/mL Lane 3: MBS150424, 0.2 ug/mL Lane 4: MBS150424,0.5 ug/mL)

Western Blot (WB)

(Loading: 15 ug of lysates per lane. Antibodies: RIP3 MBS150424,(0.5 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.)

Immunohistochemistry (IHC)

(Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-RIP3 antibody (MBS150424) at 5 ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 uC. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary.Counter stained with Hematoxylin.)

Immunohistochemistry (IHC)

(Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-RIP3 antibody (MBS150424) at 2.5ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 uC. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary.Counter stained with Hematoxylin.)

Immunofluorescence (IF)

(Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Kidney tissue labeling RIP3 with MBS150424 at 20ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).)

Western Blot (WB)

(Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in pancreas, liver spleen thymus and lung of RIP3 KO mice. Cerulein treatment upregulated RIP3 expression in the pancreas of WT mice.)

Immunohistochemistry (IHC)

(RIP3 expression detected by anti-RIP3 antibodies (MBS150424) was decreased in Fadd KO mice as compared to WT mice.)

Western Blot (WB)

(Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in RIP3 KO MEFs, but not in WT MEF cells.)

Testing Data (TD)

(Immunoblot analysis of RIP3 with anti-RIP3 antibodies (MBS150424) shows RIP3 expression was disrupted in RIP3 knockdown L929 cells.)

Testing Data (TD)

(Confocal microscopy shows colocalization of RIPK1 (green) and RIPK3 (red) in PM and RIPK3 was detected by anti-RIPK3 antibodies (MBS150424). PF or BCM suppressed LPS-induced upregulation of RIP3.)

Testing Data (TD)

(The cancer cell lines were stably expressing DYKDDDDK-tagged RIP3 and RIP expression was detected by anti-RIP3 antibodies (MBS150424) in RIP3-overexpressed cells.)

Related Product Information for anti-Ripk3 antibody

RIP3 Antibody: Certain serine/threonine protein kinases, such as ASK1, RIP, DAP, and ZIP kinases, are mediators of apoptosis. Receptor interacting proteins including RIP and RIP2/RICK mediate apoptosis induced by TNFR1 and Fas, two prototype members in the death receptor family. A novel member in the RIP kinase family was recently identified and designated RIP3. RIP3 contains N-terminal kinase domain but, unlike RIP or RIP2, lacks the C-terminal death or CARD domain. RIP3 binds to RIP and TNFR1, mediates TNFR1 induced apoptosis, and attenuates RIP and TNFR1 induced NF-κB activation. Overexpression of RIP3 induces apoptosis and NF-κB activation. The messenger RNA of RIP3 is expressed in a subset of adult tissues.

NCBI and Uniprot Product Information

• NCBI GI #: 6063101
• NCBI GeneID: 56532
• UniProt Accession #: Q9QZL0; Q3U3Z9; Q8K2Y2; G3X8V8
• Molecular Weight: Predicted: 53 kD for mouse RIP3 and 57kD for human RIP3; Observed: 53 kD for mouse RIP3 and 57kD for human RIP3
• NCBI Official Full Name: receptor interacting protein 3
• NCBI Official Synonym Full Names: receptor-interacting serine-threonine kinase 3
• NCBI Official Symbol: Ripk3
• NCBI Official Synonym Symbols: Rip3; AW107945; 2610528K09Rik
• NCBI Protein Information: receptor-interacting serine/threonine-protein kinase 3; RIP-3; mRIP3; RIP-like protein kinase 3; receptor interacting protein 3; receptor-interacting protein 3
• UniProt Protein Name: Receptor-interacting serine/threonine-protein kinase 3
• Protein Family: Receptor-interacting serine/threonine-protein kinase
• UniProt Gene Name: Ripk3
• UniProt Synonym Gene Names: Rip3; RIP-3; mRIP3
• UniProt Entry Name: RIPK3_MOUSE

Uniprot Description

RIPK3: Promotes apoptosis. Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Binds TRAF2 and RIPK1 and is recruited to the TNFR-1 signaling complex. 3 isoforms of the human protein are produced by alternative splicing.

Protein type: EC 2.7.11.1; Protein kinase, Ser/Thr (non-receptor); Protein kinase, TKL; Kinase, protein; TKL group; RIPK family

Cellular Component: mitochondrion; membrane; cytoplasm; plasma membrane; intracellular; cytosol

Molecular Function: transferase activity; NF-kappaB-inducing kinase activity; identical protein binding; protein serine/threonine kinase activity; protein binding; transferase activity, transferring phosphorus-containing groups; protein complex binding; nucleotide binding; kinase activity; ATP binding; protein kinase activity

Biological Process: spleen development; regulation of activated T cell proliferation; I-kappaB kinase/NF-kappaB cascade; viral reproduction; thymus development; protein heterooligomerization; protein amino acid autophosphorylation; T cell differentiation in the thymus; programmed cell death; positive regulation of transferase activity; lymph node development; protein amino acid phosphorylation; activation of NF-kappaB transcription factor; regulation of T cell mediated cytotoxicity; regulation of adaptive immune response; regulation of interferon-gamma production; activation of protein kinase activity; T cell homeostasis; positive regulation of ligase activity; phosphorylation; protein homooligomerization; positive regulation of oxidoreductase activity