Invitrogen

RIP3 Recombinant Polyclonal Antibody

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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RIP3 Antibody in Immunocytochemistry (ICC/IF) — For immunofluorescence analysis, HT-29 cells were fixed and permeabilized for detection of endogenous RIP3 using Anti-RIP3 Recombinant Rabbit Polyclonal Antibody (Product # 711691, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of RIP3 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of RIP3. Panel e) represents control cells with no primary antibody to assess background. Panel f) represents a negative control on HCT116 cells which are reported RIP3 negative. Images were captured at 60X magnification.

RIP3 Antibody in Western Blot (WB) — Western blot analysis was performed on membrane extracts (30 µg lysate) of HT-29 (Lane 1), THP-1 (Lane 2), Jurkat (Lane 3), Ramos (Lane 4), K-562 (Lane 5), A549 (Lane 6), MDA-MB-231 (Lane 7) and HeLa (Lane 8). The blots were probed with Anti-RIP3 Recombinant Rabbit Polyclonal Antibody (Product # 711691, 1 µg/mL). A 57 kDa band corresponding to RIP3 was observed across the cell lines tested. The blot was detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800).Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

RIP3 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Relative expression of RIP3 was observed in HT-29 and HCT 116 in Western blot using Anti-RIP3 Recombinant Rabbit Monoclonal Antibody (Product # 711691). HT-29 and HCT-116 (colorectal carcinoma) are documented to have differential basal expression of RIP-3, where-in RIP3 expression is higher in HT-29 when compared to HCT-116. {RE}

Product Details

711691

Applications	Tested Dilution	Publications
Western Blot (WB)	1 µg/mL	-
Immunocytochemistry (ICC/IF)	2 µg/mL	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Polyclonal
Type	Antibody
Immunogen	Peptide corresponding to human RIP3 (aa496-aa513) View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2688314

Product Specific Information

This antibody is predicted to react with Mouse, Rat, Horse

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

RIP3 is a novel member of RIP Ser/Thr kinase family. It is a potent inducer of apoptosis and is an important component of TNFR1 signaling complex. RIP3 consists of an N-terminal RIP-like-Kinase domain and a unique, non-homologous C-terminus responsible for caspase activation and apoptosis induction. RIP3 is expressed in multiple tissues including hematopoietic cells and plays a functional role in the regulation of apoptosis and NF-kB signaling. It is recruited to the TNFR1 signaling complex in a RIP-dependant manner where it induces apoptosis by activating caspases and/or inhibiting TNFR1-induced NF-kB activation. RIP3 negatively regulates the TLR3-Trif-mediated NF-kB signaling pathway by competing with the binding of Trif to RIP1. The human RIP3 gene is localized in the chromosomal region 14q11.2.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: receptor interacting protein 3; Receptor-interacting protein 3; receptor-interacting serine-threonine kinase 3; Receptor-interacting serine/threonine-protein kinase 3; RIP-3; RIP-like protein kinase 3

Gene Aliases: RIP3; RIPK3

UniProt ID: (Human) Q9Y572

Entrez Gene ID: (Human) 11035

Molecular Function: non-receptor serine/threonine protein kinase