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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

ELISA, Gel Supershift Assay, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Knockout Validated, Western Blot

Cited:

Block/Neutralize, Gel Supershift Assay, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Knockout Validated, Proximity Ligation Assay, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

Product Specifications

Immunogen

RIPK3/RIP3 Antibody was made to a 14 amino acid peptide near the carboxy terminus of murine RIP3. The immunogen is located within the last 50 amino acids of RIP3. Amino Acid Squence: AQFGRGRGWQPFHK

Reactivity Notes

Human, mouse and rat reactivity reported in multiple pieces of scientific literature.

Specificity

Mouse RIPK3/RIP3 Antibody has one isoform (486aa, 53 kDa). Human RIP3 has 3 isoforms, including isoform 1 (518aa, 57 kDa), isoform 2 (252aa, 28 kDa) and isoform 3 (231aa, 25 kDa). Rat RIP3 also has one isoform (478aa, 52 kDa). NBP1-77299 can detect can detect isoforms of mouse and rat as well as human isoform 1.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

53 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

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 Antibody [NBP1-77299] - Analysis of RIP3 in HeLa cell lysate with RIPK3/RIP3 antibody [NBP1-77299] at (A) 1 and (B) 2 ug/mL. Observed molecular weight ~60 kDa. Theoretical molecular weight 53 kDa.
Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 Antibody [NBP1-77299] - Mouse Cell lines. 15 ug of lysates per lane. Antibodies: [NBP1-77299], (0.5 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Observed molecular weight ~50 kDa. Theoretical molecular weight 53 kDa.
Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 AntibodyBSA Free [NBP1-77299]

Western Blot: RIPK3/RIP3 Antibody [NBP1-77299] - Analysis using RIPK3/RIP3 antibody [NBP1-77299]. L929 cell line (mouse). 20 ug whole cell lysate. Primary antibody: 1:2000. Secondary antibody: 1:20000. Western blot image submitted by a verified customer review. Observed molecular weight ~57 kDa. Theoretical molecular weight 53 kDa.

Applications for RIPK3/RIP3 Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100 - 1:2000

Immunocytochemistry/ Immunofluorescence

20 ug/ml

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

5 ug/ml

Immunoprecipitation

20 ug/mL

Western Blot

0.1-0.5 ug/ml
Application Notes
Gel supershift assay reported in scientific literature [PMID: 27721066]. Immunoprecipitation reported in scientific literature [PMID: 27861127]. RIPK3/RIP3 antibody validated for WB from a verified customer review. Knockout validation reported in scientific literature [PMID: 32246911]. Knockdown validation reported in scientific literature [PMID: 31655343]. ICC/IF, IHC, and WB reported in multiple pieces of scientific literature.

Reviewed Applications

Read 1 review rated 5 using NBP1-77299 in the following applications:

Formulation, Preparation, and Storage

Purification

Peptide affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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.

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

Entrez Gene IDs

6063101 (Mouse)

Gene Symbol

RIPK3

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 number in the search box below.

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.

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