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Rabbit Recombinant Monoclonal RIP antibody. Suitable for IP, IHC-P, WB and reacts with Mouse, Human, Rat samples. Cited in 3 publications.

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Images

Western blot - Anti-RIP antibody [EPR24883-85] (AB300617), expandable thumbnail
  • Western blot - Anti-RIP antibody [EPR24883-85] (AB300617), expandable thumbnail
  • Western blot - Anti-RIP antibody [EPR24883-85] (AB300617), expandable thumbnail
  • Western blot - Anti-RIP antibody [EPR24883-85] (AB300617), expandable thumbnail
  • Western blot - Anti-RIP antibody [EPR24883-85] (AB300617), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPFlow Cyt (Intra)ICC/IFIHC-PWB
Human
Tested
Not recommended
Not recommended
Tested
Tested
Mouse
Tested
Not recommended
Not recommended
Not recommended
Tested
Rat
Expected
Not recommended
Not recommended
Not recommended
Tested

Tested
Tested

Species

Mouse

Dilution info

1/30

Notes

-

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Rat

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Target data

Function

Serine-threonine kinase which is a key regulator of TNF-mediated apoptosis, necroptosis and inflammatory pathways (PubMed:31827280, PubMed:31827281). 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:17389591, PubMed:15310755). Required for ZBP1-induced NF-kappa-B activation in response to DNA damage (By similarity).

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal RIP antibody. Suitable for IP, IHC-P, WB and reacts with Mouse, Human, Rat samples. Cited in 3 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR24883-85

Purification technique

Affinity purification Protein A

Specificity

Not suitable for mouse and rat IHC-P.

Concentration
Loading...

Storage

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

Notes

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

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.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    The identity of the lower MW band at approximately 35 kDa is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-RIP antibody - Total protein control (ab300617) staining at 1/1000 dilution.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-RIP (phospho S166) antibody [EPR25654-166] (Anti-RIP (phospho S166) antibody [EPR25654-166] ab316923) at 1/1000 dilution

    Lane 1: Untreated 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Lane 2: Untreated 293T transfected with a human wild-type RIP expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Lane 3: 293T transfected with a human wild-type RIP expression vector containing a myc-His-tag® were treated with 100 nM Calyculin for 30 minutes, whole cell lysate at 20 µg

    Lane 4: 293T transfected with a human RIP (S166A mutation) expression vector containing a myc-His-tag® were treated with 100 nM Calyculin for 30 minutes, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 75 kDa, 36 kDa

    Exposure time: 6s

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    Anti-RIPK1 antibody [EPR24883-85] (ab300617) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab300617 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 Human RIPK1 knockout THP-1 cell line ab276121 (knockout cell lysate ab284221). 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 [EPR24883-85] (ab300617) at 1/1000 dilution

    Lane 1: Wild-type THP-1 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Raji cell lysate at 20 µg

    Secondary

    Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

    Predicted band size: 75 kDa

    Observed band size: 75 kDa

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    In Western blot, Anti-RIP (phospho S166) antibody [EPR25654-166] ab316923 was shown to bind specifically to RIP. Target of interest was observed at 75 kDa in wild-type HAP1 cell lysates (lanes 1 and 3), whereas no signal observed at this size in RIP knockout cell line lysates (lanes 2 and 4).

    The expression of RIP (phospho S166) is upregulated in response to z-VAD, TNF alpha and SM-164 treatment (PMID: 32027418, PMID: 33273695).

    The identity of the lower MW band at approximately 15 kDa is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-RIP antibody - Total protein control (ab300617) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-RIP (phospho S166) antibody [EPR25654-166] (Anti-RIP (phospho S166) antibody [EPR25654-166] ab316923) at 1/1000 dilution

    Lane 1: Untreated wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 50 µg

    Lane 2: Untreated RIP knockout HAP1 whole cell lysate at 50 µg

    Lane 3: Wild-type HAP1 treated with 20 uM z-VAD for 3.5 hours, then with 20 ng/ml TNF alpha and 100 nM SM-164 added for 3 hours whole cell lysate at 50 µg

    Lane 4: RIP knockout HAP1 treated with 20 uM z-VAD for 3.5 hours, then with 20 ng/ml TNF alpha and 100 nM SM-164 added for 3 hours whole cell lysate at 50 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 75 kDa, 36 kDa

    Exposure time: 180s

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    The expression of RIP (phospho S166) is upregulated in response to z-VAD, TNF alpha and SM-164, while it is downregulated in response to necrostain treatment (PMID: 32027418, PMID: 33273695).

    The identity of the lower MW band at approximately 35 kDa is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-RIP antibody - Total protein control (ab300617) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-RIP (phospho S166) antibody [EPR25654-166] (Anti-RIP (phospho S166) antibody [EPR25654-166] ab316923) at 1/1000 dilution

    Lane 1: Untreated HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

    Lane 2: HT-29 treated with 20 uM z-VAD for 3.5 hours, then with 20 ng/ml TNF alpha, 100 nM SM-164 and 50 uM necrostatin added for 3 hours whole cell lysate (untreated membrane) at 40 µg

    Lane 3: HT-29 treated with 20 uM z-VAD for 3.5 hours, then with 20 ng/ml TNF alpha and 100nM SM-164 added for 3 hours whole cell lysate (untreated membrane) at 40 µg

    Lane 4: HT-29 treated with 20 uM z-VAD for 3.5 hours, then with 20 ng/ml TNF alpha and 100 nM SM-164 added for 3 hours whole cell lysate (Lambda phosphatase treated membrane) at 40 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 75 kDa, 36 kDa

    Exposure time: 180s

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    All lanes: Western blot - Anti-RIP antibody [EPR24883-85] (ab300617) at 1/1000 dilution

    Lane 1: Mouse testis tissue lysate at 20 µg

    Lane 2: Rat testis tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    All lanes: Western blot - Anti-RIP antibody [EPR24883-85] (ab300617) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 3: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  • Western blot - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Western blot - Anti-RIP antibody [EPR24883-85] (ab300617)

    All lanes: Western blot - Anti-RIP antibody [EPR24883-85] (ab300617) at 1/1000 dilution

    All lanes: Mouse liver tissue lysate at 20 µg

    Secondary

    Lanes 1 - 3: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Lanes 1 - 3: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution

  • Immunohistochemistry - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Immunohistochemistry - Anti-RIP antibody [EPR24883-85] (ab300617)

    Immunohistochemical analysis of paraffin-embedded (A) Wild-type HAP1 c tissue labeling RIP with ab300617 at 1/100 (5.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on (A) Wild-type HAP1 cell pellet(B) and no staining on RIPK1 knockout HAP1 cell pellet.The section was incubated with ab300617 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunohistochemistry - Anti-RIP antibody [EPR24883-85] (ab300617), expandable thumbnail

    Immunohistochemistry - Anti-RIP antibody [EPR24883-85] (ab300617)

    Immunohistochemical analysis of paraffin-embedded Human cervical carci tissue labeling RIP with ab300617 at 1/100 (5.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human cervical carcinoma.The section was incubated with ab300617 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

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Product protocols

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