Recombinant Anti-RIP antibody [EPR4689] (ab125072)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4689] to RIP
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RIP antibody [EPR4689]
See all RIP primary antibodies -
Description
Rabbit monoclonal [EPR4689] to RIP -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human RIP aa 300-450 (internal sequence).
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Positive control
- WB: Raji, Jurkat, HeLa and 293T cell lysates
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4689 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab125072 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 75 kDa.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 75 kDa. |
Target
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Function
Essential adapter molecule for the activation of NF-kappa-B. Following different upstream signals (binding of inflammatory cytokines, stimulation of pathogen recognition receptors, or DNA damage), particular RIPK1-containing complexes are formed, initiating a limited number of cellular responses. Upon TNFA stimulation RIPK1 is recruited to a TRADD-TRAF complex initiated by TNFR1 trimerization. There, it is ubiquitinated via 'Lys-63'-link chains, inducing its association with the IKK complex, and its activation through NEMO binding of polyubiquitin chains. -
Sequence similarities
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
Contains 1 death domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsProteolytically cleaved by caspase-8 during TNF-induced apoptosis. Cleavage abolishes NF-kappa-B activation and enhances pro-apototic signaling through the TRADD-FADD interaction.
Autophosphorylated on serine and threonine residues.
Ubiquitinated by 'Lys-11'-, 'Lys-48'-, 'Lys-63'- and linear-linked type ubiquitin. 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 to the termination of the TNF- or Linear polyubiquitinated; the head-to-tail polyubiquitination is mediated by the LUBAC complex. LPS-mediated activation of NF-kappa-B. Also ubiquitinated with 'Lys-11'-linked chains. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 8737 Human
- Omim: 603453 Human
- SwissProt: Q13546 Human
- Unigene: 519842 Human
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Alternative names
- Cell death protein RIP antibody
- FLJ39204 antibody
- OTTHUMP00000039163 antibody
see all
Images
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All lanes : Anti-RIP antibody [EPR4689] (ab125072) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : RIPK1 knockout THP-1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 75 kDa
Observed band size: 76 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-RIP antibody [EPR4689] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab125072 was shown to bind specifically to RIP. A band was observed at 76 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line ab276121 (knockout cell lysate ab284210). 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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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 - 4: Merged signal (red and green). Green - ab125072 (unpurified) observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab125072 was shown to specifically react with RIP in wild-type HAP1 cells. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab125072 at a dilution of 1/1000 and ab8245 (loading control to GAPDH) at a dilution of 1/10,000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging. -
Anti-RIP antibody [EPR4689] (ab125072) at 1/5000 dilution (Purified) + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 75 kDa
Observed band size: 75 kDa -
Anti-RIP antibody [EPR4689] (ab125072) at 1/1000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 75 kDa
Observed band size: 75 kDa -
All lanes : Anti-RIP antibody [EPR4689] (ab125072) at 1/1000 dilution (unpurified)
Lane 1 : Raji cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 75 kDa
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab125072 has been referenced in 4 publications.
- Tang X et al. Sirtuin 3 induces apoptosis and necroptosis by regulating mutant p53 expression in small-cell lung cancer. Oncol Rep 43:591-600 (2020). PubMed: 31894331
- Wang X et al. Tumor Necrosis Factor (TNF) Receptor Expression Determines Keratinocyte Fate upon Stimulation with TNF-Like Weak Inducer of Apoptosis. Mediators Inflamm 2019:2945083 (2019). PubMed: 31885495
- Finger JN et al. Identification of an antibody-based immunoassay for measuring direct target binding of RIPK1 inhibitors in cells and tissues. Pharmacol Res Perspect 5:N/A (2017). ELISA . PubMed: 29226625
- Wang X et al. A20 inhibits the motility of HCC cells induced by TNF-a. Oncotarget 7:14742-54 (2016). WB . PubMed: 26909601