Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320)

Overview

  • Product name

    Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52]
    See all RIP3 primary antibodies
  • Description

    Rabbit monoclonal [EPR19403-52] to RIP3 (phospho T231 + S232)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blotmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Synthetic peptide within Mouse RIP3 aa 200-300 (phospho T231 + S232). The exact sequence is proprietary. Two peptide immunogens within mouse RIP3 aa 200-300 were used for co-immunization.
    Database link: Q9QZL0

  • Positive control

    • WB: L-929 treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hr, whole cell lysate.
  • 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 ab222320 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
WB 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 57 kDa).
Dot blot 1/1000.

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 T231 + S232) antibody [EPR19403-52] (ab222320) at 1/500 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 10 ng/ml TNF-a and 20 µM z-VAD for 2 hours
    Lane 3 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 10 ng/ml TNF-a and 20 µM z-VAD for 3.5 hours
    Lane 4 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours
    Lane 5 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate with RIP3 KO cells treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

    Predicted band size: 57 kDa


    Exposure time: 60 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Cells were lysed in 4% SDS buffer (4% SDS,10% 2-ME, 20% glycerol, 0.004% Bromophenol blue, Tris-HCl).

    The images were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.

  • All lanes : Anti-RIP3 (phospho T231 + S232) antibody [EPR19403-52] (ab222320) at 1/1000 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 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours
    Lane 3 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hours, then treated with alkaline phosphatase for 1 hour

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 57 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The higher MW bands in Lanes 1 and 2 are extra bands.

    The lysates were prepared following the 1% SDS Hot Lysate buffer method.

    For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

  • Dot blot analysis of RIP3 (phospho T231 + S232) labeled with ab222320 at 1/1000 dilution.

    Lane 1: RIP3 (phospho T231 + S232) peptide (aa229-239);

    Lane 2: RIP3 (phospho T231 + S232) peptide (aa223-236);

    Lane 3: RIP3 (phospho T231) peptide (aa223-239);

    Lane 4: RIP3 (phospho S232) peptide (aa223-239);

    Lane 5: RIP3 non-phospho peptide (aa223-239).

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

References

ab222320 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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