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  1. Link

    rip3-phospho-s232-antibody-epr9516n-25-ab195117.pdf

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Cell Biology Apoptosis Receptors Associated Proteins
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RecombinantRabMAb

Recombinant Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (13) Submit a question References (42)

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Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
  • Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
  • Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
  • ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
  • Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9516(N)-25] to RIP3 (phospho S232)
  • Suitable for: ELISA, WB, Dot blot
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Carrier Free

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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. This information is proprietary to Abcam and/or its suppliers.

  • 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR9516(N)-25
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Receptors
    • Associated Proteins
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Kinases
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Associated Proteins
    • Cancer
    • Cell Death
    • Necroptosis

Associated products

  • Alternative Versions

    • Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] - BSA and Azide free (ab240336)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human RIP3 protein (Tagged) (ab125566)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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 (1)
Use at an assay dependent concentration.
WB (8)
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
Dot blot
Use at an assay dependent concentration.
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.
  • Target information above from: UniProt accession Q9Y572 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 56532 Mouse
    • SwissProt: Q9QZL0 Mouse
    • Unigene: 46612 Mouse
    • 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

    • Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      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.

    • Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      Western blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      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

    • Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      Dot Blot - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)

      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.

    • ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      ELISA - Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)

      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.

    • Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)
      Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25] (ab195117)

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (42)

    Publishing research using ab195117? Please let us know so that we can cite the reference in this datasheet.

    ab195117 has been referenced in 42 publications.

    • Wang W  et al. Inhibition of adenosine kinase attenuates myocardial ischaemia/reperfusion injury. J Cell Mol Med 25:2931-2943 (2021). PubMed: 33523568
    • Ye L  et al. CB1R Promotes Chronic Alcohol-Induced Neuronal Necroptosis in Mice Prefrontal Cortex. Alcohol Alcohol 56:230-239 (2021). PubMed: 33382401
    • Moujalled D  et al. The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis. Cell Death Dis 12:133 (2021). PubMed: 33510145
    • Yin Z  et al. RIPK1 is a negative mediator in Aquaporin 1-driven triple-negative breast carcinoma progression and metastasis. NPJ Breast Cancer 7:53 (2021). PubMed: 33980862
    • Hu Y  et al. Precision control of mTORC1 is crucial for the maintenance and IL-13 responsiveness of alveolar macrophages. Int Immunopharmacol 95:107552 (2021). PubMed: 33743315
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (rat primary cortical neuronal cultures)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Treatment
    shikonin 20uM for 1h
    Specification
    rat primary cortical neuronal cultures
    Blocking step
    casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 17 2021

    Western blot abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - other (HEK293T)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    10 µg
    Specification
    HEK293T
    Blocking step
    Serum as blocking agent for 1 hour(s) and 20 minute(s) · Concentration: 10% · Temperature: 30°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 20 2021

    ELISA abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Mouse Serum (Blood)
    Specification
    Blood
    Blocking step
    Milk as blocking agent for 1 hour(s) and 20 minute(s) · Concentration: 5% · Temperature: 37°C
    Type
    Sandwich (Detection)
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 20 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Embryo)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
    Permeabilization
    Yes - 0.3% Triton X-100
    Specification
    Embryo
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 30 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Pancreas)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    No
    Specification
    Pancreas
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 11 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 18 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 26 2017

    Western blot abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 21 2016

    Western blot abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Miss. Berta Anuncibay

    Verified customer

    Submitted Oct 10 2016

    Western blot abreview for Anti-RIP3 (phospho S232) antibody [EPR9516(N)-25]

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 11 2016

    1-10 of 13 Abreviews or Q&A

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