Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse (Negative), Primate

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot, Proximity Ligation Assay

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all RIPK3/RIP3 Primary Antibodies »

Product Specifications

Immunogen

RIPK3/RIP3 Antibody was made to a synthetic peptide (QEGPKDPEAWSRPQ) corresponding to amino acids 497-518 of human RIPK3/RIP3 was used as immunogen for this antibody (isoform CRA_a, 518 amino acids, NP_006862.2). This also corresponds to amino acids 298-311 of isoform CRA_b (GenBank: EAW66022.1).

Reactivity Notes

Chimpanzee (100%). This antibody is not suitable for testing of mouse RIP3. Human reactivity reported in multiple pieces of scientific literature.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

56.7 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 - BSA Free

Immunohistochemistry-Paraffin: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588]

Immunohistochemistry-Paraffin: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588]

Immunohistochemistry-Paraffin: RIPK3/RIP3 Antibody [NBP2-24588] - Analysis of RIP3 using polyclonal RIPK3/RIP3 Antibody [NBP2-24588]. Human pancreas probed with RIPK3/RIP3 antibody at 10 ug/ml.
Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP2-24588]

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP2-24588]

RIPK3-RIP3-Antibody-Western-Blot-NBP2-24588-img0006.jpg
Immunocytochemistry/ Immunofluorescence: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588]

Immunocytochemistry/ Immunofluorescence: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588]

Immunocytochemistry/Immunofluorescence: RIPK3/RIP3 Antibody [NBP2-24588] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with RIPK3/RIP3 Antibody [NBP2-24588] at a 1:200 dilution overnight at 4C and detected with DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used at a 1:1000 dilution as a co-stain and detected with DyLight 550 (Red) at a 1:500 dilution. Nuclei were detected with DAPI (Blue) at 2.0ug/ml in 1X PBS. Cells were imaged using a 40X objective.
Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP2-24588]

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP2-24588]

Western Blot: RIPK3/RIP3 Antibody [NBP2-24588] - Analysis of RIP3 using polyclonal RIPK3/RIP3 antibody [NBP2-24588]. Human tissue blot probed with 5 ug/ml of RIP3 antibody: 1) brain, 2) heart, 3) small intestine, 4) kidney, 5) liver, 6) lung, 7) skeletal muscle, 8) stomach, 9) spleen, 10) ovary, 11) testis, 12) placenta probed at 5 ug/ml, 13) Ramos cell lysate probed at 7 ug/ml. Theoretical molecular weight 56.7 kDa.
RIPK3/RIP3 Antibody - BSA Free

Western Blot: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588] -

Western Blot: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588] - HOIP limits the activity of TRAIL‐induced apoptosis‐ & necroptosis‐mediating signalling complexesControl & HOIP KO HeLa cells were treated with FLAG‐lz‐TRAIL (500 ng/ml) for the indicated times. The TRAIL complex I was immunoprecipitated via anti‐FLAG beads. Complex II was isolated by immunoprecipitating caspase‐8 from complex I‐depleted lysates. Western blot was performed using the indicated antibodies. * indicates unspecific bands.WT & HOIP‐deficient TNF KO MEFs, pre‐treated for 1 h with zVAD, were stimulated with iz‐TRAIL (1 μg/ml) for the indicated times. FADD‐containing complexes were immunoprecipitated & analysed by Western blot.Data information: See also Fig EV2. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28258062), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RIPK3/RIP3 Antibody - BSA Free

Western Blot: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588] -

Western Blot: RIPK3/RIP3 Antibody - BSA Free [NBP2-24588] - The catalytic activity of HOIP contributes to preventing TRAIL‐induced apoptosis but is dispensable for preventing necroptosisLysates from control or HOIP KO K562 cells reconstituted w/ empty vector, HOIP WT or HOIP C885S analysed by WB.HOIP KO K562 cells reconstituted w/ empty vector, HOIP WT or HOIP C885S, pre‐treated w/ zVAD and/or Nec‐1s for 1 h as indicated, treated w/ iz‐TRAIL (1 μg/ml) for 24 h. Percentage of cell death determined by flow cytometry after PI labelling (n = 5; mean ± SEM). *P < 0.05, **P < 0.01; statistics performed using ANOVA.Control & HOIP KO K562 cells reconstituted w/ empty vector, HOIP WT or HOIP C885S treated w/ iz‐TRAIL (100 ng/ml) for the indicated times. Lysates analysed w/ the indicated antibodies.Lysates from TNF KO HOIP KO MEFs reconstituted w/ empty vector, HOIP WT or HOIP C885S analysed by WB.TNF KO HOIP KO MEFs reconstituted w/ empty vector, HOIP WT or HOIP C885S, pre‐treated w/ zVAD & Nec‐1s for 1 h as indicated, treated w/ iz‐TRAIL (1 μg/ml) for 24 h. Percentage of cell death determined by flow cytometry after PI labelling (n = 4; mean ± SEM). *P < 0.05, ***P < 0.001; statistics performed using ANOVA.TNF KO HOIP KO MEFs reconstituted w/ empty vector, pre‐treated w/ zVAD and/or the RIPK3 inhibitor GSK'872 for 1 h as indicated, treated w/ iz‐TRAIL (1 μg/ml) for 24 h. Percentage of cell death determined by flow cytometry after PI labelling (n = 3; mean ± SEM). *P < 0.05; statistics performed using t‐test.TNF KO HOIP KO MEFs reconstituted w/ empty vector, HOIP WT or HOIP C885S, pre‐treated w/ zVAD for 1 h, treated w/ iz‐TRAIL (1 μg/ml) for the indicated times. FADD‐containing complexes immunoprecipitated & analysed by WB. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28258062), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for RIPK3/RIP3 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:20-1:500

Immunohistochemistry-Paraffin

10 ug/ml

Western Blot

3-7 ug/ml
Application Notes
Use in Immunoprecipitation reported in scientific literature. Use in Western Blot reported in multiple pieces of scientific literature.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at -20C. 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

UniProt

Q9Y572 (Human)

Additional RIPK3/RIP3 Products

Product Documents for RIPK3/RIP3 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for RIPK3/RIP3 Antibody - BSA Free

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

Citations for RIPK3/RIP3 Antibody - BSA Free

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for RIPK3/RIP3 Antibody - BSA Free

Showing  1 - 5 of 5 FAQs Showing All

  • 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: The customer would like to learn more about the IHC image of shown within the webpage. Was this figure a result from frozen sections or formalin/PFA-fixed paraffin-embedded sections? Or perhaps some other section? They would also would like to have the recommended protocol, for details especially regarding pH value. Would you please help confirming this?

    A: Our RIPK3 antibody with catalogue number NBP2-24588 (also known as IMG-5846A) has been validated for Western blotting and for IHC with paraffin-embedded samples. It has not yet been tested for IHC with frozen tissue. The IHC image on our website shows human pancreas tissue probed with NBP2-24588 at a concentration of 10ug/ml. Please find here links to our protocol for IHC-P, and also our antigen retrieval protocol: IHC-P and Antigen retrieval.  For antigen retrieval we recommend the use of citrate buffer.

  • 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.

  • Q: Would you please confirm the concentration for item NBP2-24588, Lot A-1?

    A: The concentration is 1 mg/ml.

  • 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: The customer would like to learn more about the IHC image of shown within the webpage. Was this figure a result from frozen sections or formalin/PFA-fixed paraffin-embedded sections? Or perhaps some other section? They would also would like to have the recommended protocol, for details especially regarding pH value. Would you please help confirming this?

    A: Our RIPK3 antibody with catalogue number NBP2-24588 (also known as IMG-5846A) has been validated for Western blotting and for IHC with paraffin-embedded samples. It has not yet been tested for IHC with frozen tissue. The IHC image on our website shows human pancreas tissue probed with NBP2-24588 at a concentration of 10ug/ml. Please find here links to our protocol for IHC-P, and also our antigen retrieval protocol: IHC-P and Antigen retrieval.  For antigen retrieval we recommend the use of citrate buffer.

  • 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.

  • Q: Would you please confirm the concentration for item NBP2-24588, Lot A-1?

    A: The concentration is 1 mg/ml.

  • 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: The customer would like to learn more about the IHC image of shown within the webpage. Was this figure a result from frozen sections or formalin/PFA-fixed paraffin-embedded sections? Or perhaps some other section? They would also would like to have the recommended protocol, for details especially regarding pH value. Would you please help confirming this?

    A: Our RIPK3 antibody with catalogue number NBP2-24588 (also known as IMG-5846A) has been validated for Western blotting and for IHC with paraffin-embedded samples. It has not yet been tested for IHC with frozen tissue. The IHC image on our website shows human pancreas tissue probed with NBP2-24588 at a concentration of 10ug/ml. Please find here links to our protocol for IHC-P, and also our antigen retrieval protocol: IHC-P and Antigen retrieval.  For antigen retrieval we recommend the use of citrate buffer.

  • 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.

  • Q: Would you please confirm the concentration for item NBP2-24588, Lot A-1?

    A: The concentration is 1 mg/ml.

  • 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: The customer would like to learn more about the IHC image of shown within the webpage. Was this figure a result from frozen sections or formalin/PFA-fixed paraffin-embedded sections? Or perhaps some other section? They would also would like to have the recommended protocol, for details especially regarding pH value. Would you please help confirming this?

    A: Our RIPK3 antibody with catalogue number NBP2-24588 (also known as IMG-5846A) has been validated for Western blotting and for IHC with paraffin-embedded samples. It has not yet been tested for IHC with frozen tissue. The IHC image on our website shows human pancreas tissue probed with NBP2-24588 at a concentration of 10ug/ml. Please find here links to our protocol for IHC-P, and also our antigen retrieval protocol: IHC-P and Antigen retrieval.  For antigen retrieval we recommend the use of citrate buffer.

  • 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.

  • Q: Would you please confirm the concentration for item NBP2-24588, Lot A-1?

    A: The concentration is 1 mg/ml.

  • 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: The customer would like to learn more about the IHC image of shown within the webpage. Was this figure a result from frozen sections or formalin/PFA-fixed paraffin-embedded sections? Or perhaps some other section? They would also would like to have the recommended protocol, for details especially regarding pH value. Would you please help confirming this?

    A: Our RIPK3 antibody with catalogue number NBP2-24588 (also known as IMG-5846A) has been validated for Western blotting and for IHC with paraffin-embedded samples. It has not yet been tested for IHC with frozen tissue. The IHC image on our website shows human pancreas tissue probed with NBP2-24588 at a concentration of 10ug/ml. Please find here links to our protocol for IHC-P, and also our antigen retrieval protocol: IHC-P and Antigen retrieval.  For antigen retrieval we recommend the use of citrate buffer.

  • 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.

  • Q: Would you please confirm the concentration for item NBP2-24588, Lot A-1?

    A: The concentration is 1 mg/ml.

Showing  1 - 5 of 5 FAQs Showing All
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