AB178420

Anti-RIP antibody [EPR4689-100]

20ul selling size
KO Validated
RabMAb
Recombinant
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(2 Publications)

Knockout Tested Rabbit Recombinant Monoclonal RIP antibody. Suitable for WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.

View Alternative Names

RIP, RIP1, RIPK1, Receptor-interacting serine/threonine-protein kinase 1, Cell death protein RIP, Receptor-interacting protein 1, RIP-1

5 Images

Flow Cytometry (Intracellular) - Anti-RIP antibody [EPR4689-100] (AB178420)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RIP antibody [EPR4689-100] (AB178420)

Intracellular flow cytometric analysis of permeabilized HeLa cells labeling RIP with ab178420 at 1/10 dilution (red) compared with a rabbit IgG negative control (green).

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Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : RIP knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : Raji cell lysate (20 μg)
Lanes 1 to 4 : Merged signal (red and green). Green - ab178420 observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab178420 was shown to react with RIP in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab178420 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)

Predicted band size: 75 kDa

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Lab

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Human RIPK1 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-ripk1-knockout-thp-1-cell-line-ab276121'>ab276121</a>)

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 75 kDa

Observed band size: 76 kDa

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Lab

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

Western blot : Anti-RIPK1 antibody [EPR4689-100] (ab178420) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab178420 was shown to bind specifically to RIPK1. A band was observed at 75 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line. To generate this image, wild-type and RIPK1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

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Supplier Data

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

HeLa cell lysate with Staurosporine at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 75 kDa

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Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR4689-100

Isotype

IgG

Carrier free

Reacts with

Human

Applications

WB, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000 - 1/10000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/10 - 1/100", "FlowCytIntra-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody." } } }

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Product details

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 50% Tissue culture supernatant, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

RIP also known as Receptor-Interacting Protein or RIPK1 is a serine/threonine-protein kinase with a mass of approximately 74 kDa. It plays an important role in cell death and survival signaling pathways. RIP is expressed ubiquitously across various tissues indicating its importance in many cellular functions. The protein contains a kinase domain an intermediate domain for protein-protein interactions and a death domain which facilitates its involvement in apoptotic signaling processes.

Biological function summary

Receptor-Interacting Protein Kinase 1 (RIPK1) participates in regulating both necroptosis and apoptosis distinguishing itself as an important mediator in cell death mechanisms. As part of the necrosome complex which includes RIPK3 and MLKL RIPK1 functions in necroptosis—a programmed form of necrosis. This characteristic involvement shows its dual role in maintaining cell fate decisions making it an integral part of immune response and inflammation control.

Pathways

RIPK1 strongly associates with the TNF signaling pathway and NF-kB pathway. Its interaction with TNF receptor 1 (TNFR1) and consequent involvement with TRADD and TRAF2 mediates the signal transduction necessary for the activation of NF-kB leading to transcription of genes involved in survival and inflammation. This connection illustrates its capability to switch between promoting cell survival through NF-kB and facilitating cell death via necroptosis or apoptosis depending on cellular context and cues.

RIPK1 plays a significant role in conditions such as inflammatory diseases and neurodegenerative disorders. Its overactivation results in excessive cell death implicated in inflammatory conditions; necrostatin a necroptosis inhibitor targets RIPK1 to potentially mitigate this damage. Furthermore RIPK1's dysregulation links to Alzheimer's disease where it can interact with components like RIPK3 to exacerbate neurodegenerative processes. This relationship underlines the potential of targeting RIPK1 therapeutically to manage inflammation and neurodegeneration.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Serine-threonine kinase which is a key regulator of TNF-mediated apoptosis, necroptosis and inflammatory pathways (PubMed : 17703191, PubMed : 24144979, PubMed : 31827280, PubMed : 31827281, PubMed : 32657447, PubMed : 35831301). Exhibits kinase activity-dependent functions that regulate cell death and kinase-independent scaffold functions regulating inflammatory signaling and cell survival (PubMed : 11101870, PubMed : 19524512, PubMed : 19524513, PubMed : 29440439, PubMed : 30988283). Has kinase-independent scaffold functions : 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-kappa-B pathway (By similarity). Kinase activity is essential to regulate necroptosis and apoptosis, two parallel forms of cell death : upon activation of its protein kinase activity, regulates assembly of two death-inducing complexes, namely complex IIa (RIPK1-FADD-CASP8), which drives apoptosis, and the complex IIb (RIPK1-RIPK3-MLKL), which drives necroptosis (By similarity). RIPK1 is required to limit CASP8-dependent TNFR1-induced apoptosis (By similarity). In normal conditions, RIPK1 acts as an inhibitor of RIPK3-dependent necroptosis, a process mediated by RIPK3 component of complex IIb, which catalyzes phosphorylation of MLKL upon induction by ZBP1 (PubMed : 19524512, PubMed : 19524513, PubMed : 29440439, PubMed : 30988283). Inhibits RIPK3-mediated necroptosis via FADD-mediated recruitment of CASP8, which cleaves RIPK1 and limits TNF-induced necroptosis (PubMed : 19524512, PubMed : 19524513, PubMed : 29440439, PubMed : 30988283). Required to inhibit apoptosis and necroptosis during embryonic development : acts by preventing the interaction of TRADD with FADD thereby limiting aberrant activation of CASP8 (By similarity). In addition to apoptosis and necroptosis, also involved in inflammatory response by promoting transcriptional production of pro-inflammatory cytokines, such as interleukin-6 (IL6) (PubMed : 31827280, PubMed : 31827281). Phosphorylates RIPK3 : RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation (PubMed : 19524513). Phosphorylates DAB2IP at 'Ser-728' in a TNF-alpha-dependent manner, and thereby activates the MAP3K5-JNK apoptotic cascade (PubMed : 15310755, PubMed : 17389591). Required for ZBP1-induced NF-kappa-B activation in response to DNA damage (By similarity).

See full target information RIPK1

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Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Science signaling 13: PubMed32398349

2020

The death-inducing activity of RIPK1 is regulated by the pH environment.

Applications

Unspecified application

Species

Unspecified reactive species

Kenta Moriwaki,Sakthi Balaji,Francis Ka-Ming Chan

Carcinogenesis 38:402-410 PubMed28334272

2017

17β-estradiol-induced ACSL4 protein expression promotes an invasive phenotype in estrogen receptor positive mammary carcinoma cells.

Applications

Unspecified application

Species

Unspecified reactive species

Anissa Belkaid,Rodney J Ouellette,Marc E Surette

View all publications

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