Q: If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?

A: If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product. Please check out our Innovator's Reward Program if you decide to test an antibody with a species or application that is not currently listed.

Q: What is the theoretical molecular weight for your RIPK3/RIP3 antibodies?

A: This depends on the immunogen. We have one RIPK3/RIP3 antibody [NBP1-77299] that is raised against murine RIP3. The theoretical molecular weight for this product is 53 kDa. The rest of our RIPK3/RIP3 antibodies are developed against human RIP3 and have a theoretical molecular weight of 56.7 kDa.

Q: What research areas can this product be used in?

A: All RIPK3/RIP3 products can be used in: Cancer, Cell Biology, Immunology, Necroptosis, Protein Kinase, Signal Transduction.

RIPK3/RIP3 Antibody (PSH04-78)

Novus Biologicals | Catalog # NBP3-32905

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Product Documents

Recombinant Monoclonal Antibody

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH04-78 expressed in HEK293

View all RIPK3/RIP3 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide within human RIPK3/RIP3 aa 401-450 / 518. (Uniprot: Q9Y572)

Localization

Cytoplasm, cytosol, Nucleus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

57 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for RIPK3/RIP3 Antibody (PSH04-78)

Western Blot: RIPK3/RIP3 Antibody (PSH04-78) [NBP3-32905]

Western Blot: RIPK3/RIP3 Antibody (PSH04-78) [NBP3-32905] - Western blot analysis of RIPK3/RIP3 on different lysates with Rabbit anti-RIPK3/RIP3 antibody (NBP3-32905) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: HT-29 cell lysate
Lane 3: SK-Br-3 cell lysate
Lane 4: HeLa cell lysate (negative)

Lysates/proteins at 30 ug/Lane.

Predicted band size: 57 kDa
Observed band size: 55 kDa

Exposure time: 1 minute; ECL;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32905) at 1/2,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: RIPK3/RIP3 Antibody (PSH04-78) [NBP3-32905]

Immunohistochemistry: RIPK3/RIP3 Antibody (PSH04-78) [NBP3-32905] - Immunohistochemical analysis of paraffin-embedded HT-29 (positive) and HeLa (negative) cells with Rabbit anti-RIPK3/RIP3 antibody (NBP3-32905) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32905) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Applications for RIPK3/RIP3 Antibody (PSH04-78)

Application

Recommended Usage

Immunohistochemistry-Paraffin

1:2000-1:5000

Western Blot

1:2000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: RIPK3/RIP3

RIPK3, Receptor interacting serine/threonine kinase 3 or RIP-like protein kinase 3 (human RIPK3 isoform1 theoretical molecular weight 57kDa) is a cytosolic protein with an amino terminal active kinase domain. RIP kinases, RIPK1 and RIPK3 play a central role in the induction of necroptosis, a form of programmed and inflammatory cell death that is caspase-independent (1). Necroptosis is a form of programmed necrosis which is initiated through the activation of RIPK3 by various ligands such as Fas, LPS and TNF. Formation of the necrosome results from the interaction between RIPK1 and RIPK3 mediated through their RIP homotypic interaction motifs (RHIMs) (1, 2). Following necrosome formation, RIPK3 activation leads to the phosphorylation of mixed lineage kinase domain-like protein (MLKL) which induces its oligomerization and translocation to the cell membrane where it alters plasma membrane integrity (2). Loss of membrane integrity results in lytic cell death, release of damage associated molecular patterns (DAMPs) and inflammation (2, 3). Therefore, the interaction of RIPK3 with other RHIM proteins is necessary for the initiation of necroptosis, while the execution phase is dependent on the activation of MLKL (4).

RIPK3 has several phosphorylation sites that are required for its role in necroptosis. For example, serine204 in mouse, which is conserved in the human (serine199), is necessary for necroptosis while serine residues 232 and 227 are both required for RIPK3s interaction with MLKL (2). The core necroptosis proteins RIPK1/RIPK3 and MLKL are implicated in several disease states such as neurodegeneration, cardiovascular, hepatic and pulmonary disease (3).

References

1. Orozco, S., & Oberst, A. (2017). RIPK3 in cell death and inflammation: the good, the bad, and the ugly. Immunological Reviews. https://doi.org/10.1111/imr.12536

2. Dhuriya, Y. K., & Sharma, D. (2018). Necroptosis: A regulated inflammatory mode of cell death. Journal of Neuroinflammation. https://doi.org/10.1186/s12974-018-1235-0

3. Choi, M. E., Price, D. R., Ryter, S. W., & Choi, A. M. K. (2019). Necroptosis: A crucial pathogenic mediator of human disease. JCI Insight. https://doi.org/10.1172/jci.insight.128834

4. Lee, K.-H., & Kang, T.-B. (2019). The Molecular Links between Cell Death and Inflammasome. Cells. https://doi.org/10.3390/cells8091057

Long Name

Receptor (TNFRSF)-Interacting Serine-Threonine Kinase 3

Alternate Names

RIP3

Gene Symbol

RIPK3

Additional RIPK3/RIP3 Products

Product Documents for RIPK3/RIP3 Antibody (PSH04-78)

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Certificate of Analysis

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Product Specific Notices for RIPK3/RIP3 Antibody (PSH04-78)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Apoptosis Adaptor Proteins

NFkB Pathway

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