Overview

  • Product name
    Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]
    See all RIP3 primary antibodies
  • Description
    Rabbit monoclonal [EPR9516(N)-25] to RIP3 (phospho S232)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ELISA, WB, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Mouse RIP3 aa 200-300 (phospho S232). The exact sequence is proprietary.
    Database link: Q9QZL0

  • Positive control
    • WB: L-929 whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 8 h and then harvested; Untreated L-929 whole cell lysate; TSE (WT) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested; TSE (KO MLKL) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab195117 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
Dot blot Use at an assay dependent concentration.

Target

  • Function
    Promotes apoptosis.
  • Tissue specificity
    Detected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.
  • Sequence similarities
    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Autophosphorylated.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Receptor interacting protein 3 antibody
    • Receptor interacting serine threonine kinase 3 antibody
    • Receptor interacting serine/threonine protein kinase 3 antibody
    • Receptor-interacting protein 3 antibody
    • Receptor-interacting serine/threonine-protein kinase 3 antibody
    • RIP 3 antibody
    • RIP like protein kinase 3 antibody
    • RIP-3 antibody
    • RIP-like protein kinase 3 antibody
    • RIPK 3 antibody
    • RIPK3 antibody
    • RIPK3_HUMAN antibody
    see all

Images

  • All lanes : Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117) at 1/10000 dilution

    Lane 1 : Untreated L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
    Lane 2 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate treated with phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117) at 1/2000 dilution

    Lane 1 : Untreated TSE (WT) whole cell lysate
    Lane 2 : TSE (WT) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested
    Lane 3 : Untreated TSE (KO RIP3) whole cell lysate
    Lane 4 : TSE (KO RIP3) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested
    Lane 5 : Untreated TSE (KO MLKL) whole cell lysate
    Lane 6 : TSE (KO MLKL) whole cell lysate treated with 20 ng/ml TNF alpha,100 nM Smac mimetic, and 20 µM z-VAD for 12 h and then harvested

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Lysates were provided by Xiaodong Wang, National Institute of Biological Sciences, Beijing

  • Primary antibody: ab195117 at 1:1000 dilution (1.6μg/ml), secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:1000 dilution, Blocking and dilution buffer: 5% NFDM/TBST, Lane 1: RIP3 (phospho S232) phospho peptide, Lane 2: RIP3 Non-phospho peptide, Exposure time: 3 minutes.

    We observed weak binding to the non-phospho peptide at very high concentrations.

  • Serially diluted ab195117 was bound to immobilized phospho or non-phospho (control) peptide demonstrating minimal cross-reactivity to the non-phosphorylated residue. Antigen - RIP3 (phospho S232) phospho peptide;RIP3 non-phospho peptide at 250 ng/ml. Primary antibody concentration range: 0 - 500 ng/ml. Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1:2500.

References

This product has been referenced in:
  • Zechendorf E  et al. Heparan Sulfate Induces Necroptosis in Murine Cardiomyocytes: A Medical-In silico Approach Combining In vitro Experiments and Machine Learning. Front Immunol 9:393 (2018). WB ; Mouse . Read more (PubMed: 29616016) »
  • Fuchslocher Chico J  et al. The enhanced susceptibility of ADAM-17 hypomorphic mice to DSS-induced colitis is not ameliorated by loss of RIPK3, revealing an unexpected function of ADAM-17 in necroptosis. Oncotarget 9:12941-12958 (2018). Read more (PubMed: 29560122) »
See all 8 Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 18 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary hippocampal neurons treated with TNF-a/ZVA)
Permeabilization
Yes - triton X-100
Specification
Primary hippocampal neurons treated with TNF-a/ZVA
Blocking step
Gelatin from cold water fish skin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 26 2017

Application
Western blot
Sample
Rabbit Cell lysate - whole cell (Rabbit Reticulocyte)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
0.5 µg
Specification
Rabbit Reticulocyte
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 21 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
40 µg
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Miss. Berta Anuncibay

Verified customer

Submitted Oct 10 2016

Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (VERO E6)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
50000 cells
Treatment
TNFalpha (25ng/mL) and zVAD (50uM) for 5 hours
Specification
VERO E6
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 11 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
50000 cells
Treatment
TNFalpha (25ng/mL) and zVAD (50uM) for 6 hours
Specification
BHK-21 (Kidney Fibroblasts)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 19 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Human Foreskin Fibroblasts)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
50000 cells
Treatment
Infected with human cytomegalovirus for 72 hrs
Specification
Human Foreskin Fibroblasts
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 02 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
75000 cells
Treatment
TNFalpha (50ng/mL) and zVAD (50uM) for 12 hours
Specification
Mouse embryonic fibroblasts (MEFs)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Apr 02 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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