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Purified anti-RIP-1 Antibody

Pricing & Availability
Clone
W17101A (See other available formats)
Regulatory Status
RUO
Other Names
Receptor-Interacting Protein Kinase 1, RIP, RIP-1, RIP1, IMD57, AIEFL, Receptor-Interacting Serine/Threonine-Protein Kinase
Isotype
Rat IgG2b, λ
Ave. Rating
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Product Citations
publications
W17101A_PURE_RIP-1_Antibody_1_033122
Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.5 μg/mL (1:1000 dilution) purified anti-RIP-1 (clone W17101A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG (Cat. No. 405405) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:25000 dilution. Lane M: Molecular weight marker
  • W17101A_PURE_RIP-1_Antibody_1_033122
    Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.5 μg/mL (1:1000 dilution) purified anti-RIP-1 (clone W17101A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG (Cat. No. 405405) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:25000 dilution. Lane M: Molecular weight marker
  • W17101A_PURE_RIP-1_Antibody_2_033122
    ICC staining of purified anti-RIP-1 (clone W17101A) on HeLa cells. The cells were fixed and permeabilized with ice-cold methanol and blocked with 5% FBS for 1 hour at room temperature. The cells were then stained with 5.0 µg/mL of the primary antibody, followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 goat anti-rat IgG (Cat. No. 405422) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801), and the image was captured with a 60X objective. Scale bar: 10 µm
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603351 25 µg $118
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603352 100 µg $293
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Description

Receptor-interacting serine/threonine-protein kinase 1 (RIP-1) is a member of the RIP family of serine/threonine kinases and consists of a kinase domain (KD) at its N-terminus, and a death domain (DD) in its C-terminus. It is a key regulator of cellular inflammatory response and cell survival when functioning as a scaffold protein, namely as part of the NF-κB signaling pathway, and is a key regulator of cell death when functioning as a kinase. RIP-1 promotes cell survival as part of the tumor necrosis factor receptor 1 (TNFR1) complex following ubiquitination by M1 and K63, and phosphorylation mediated by IKK. When ubiquitination is disrupted by ubiquitin E3 ligases, RIP-1 associates with FADD and caspase 8 to promote apoptosis or forms a complex with RIPK3 to induce necroptosis. Due to it being part of the TNF signaling pathway and its dual roles in cell survival and cell death, RIP-1 is of particular interest as a therapeutic target, namely as a mediator in the induction of immunogenic cell death and the treatment of cancer as well as in the treatment of autoimmune and neurodegenerative disorders.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human RIP1 protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.5 µg/mL. For immunocytochemistry, a concentration of 5 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using HeLa cells with localization expected in the cytosol and plasma membrane. Three fix/perm methods were used: methanol fixation, or 4% PFA fixation followed by permeabilization with either methanol or Triton X-100. Methanol-only fixation produced a strong signal with the expected cytoplasmic localization. PFA-fixation followed by permeabilization with either methanol or Triton X-100 did not produce the expected cytoplasmic/membrane signal.

This clone was tested for IHC-P on human skin tissue using Biolegend’s AEC Chromagen. Positive signal was not observed.

RRID
AB_2922634 (BioLegend Cat. No. 603351)
AB_2922634 (BioLegend Cat. No. 603352)

Antigen Details

Structure
RIP-1 is a 671 amino acid protein with a predicted molecular weight of 76 kD.
Distribution

Ubiquitously expressed/localized to the cell membrane and cytoplasm

Function
Serine/threonine-protein kinase, Transferase
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Death, Neurodegeneration, Neuroscience
Molecular Family
Protein Kinases/Phosphatase
Antigen References
  1. Ang RL, et al. 2019. Front Cell Dev Biol. 7:163.
  2. Degterev A, et al. 2019. Proc Natl Acad Sci U S A. 116:9714-22.
  3. Moriwaki K, et al. 2021. Mucosal Immunol. 15:84-95.
  4. Smith HG, et al. 2020. EMBO Mol Med. 12:e10979.
Gene ID
8737 View all products for this Gene ID
UniProt
View information about RIP-1 on UniProt.org
Go To Top Version: 1    Revision Date: 03/31/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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