Anti-RIP3 antibody (ab56164)

Rabbit polyclonal RIP3 antibody. Validated in WB, IP, IHC and tested in Mouse, Human. Cited in 23 publication(s). Independently reviewed in 26 review(s). Immunogen corresponding to synthetic peptide.

Overview

  • Product name
  • Description
    Rabbit polyclonal to RIP3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human RIP3 aa 168-217. Immunogen is located within the provided sequence.
    Sequence:

    GGSQSGTGSGEPGGTLGYLAPELFVNVNRKASTASDVYSFGILMWAVLAG


    Database link: Q9Y572
    (Peptide available as ab178834)

  • Positive control
    • Jurkat cell lysate (Western blot). Human pancreas (IHC-P).

Properties

Applications

Our Abpromise guarantee covers the use of ab56164 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
IP Use at an assay dependent concentration.
WB Use a concentration of 1.25 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 3 - 12 µg/ml.

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

  • Predicted band size: 57 kDa
    Observed band size: 57 kDa

  • All lanes : Anti-RIP3 antibody (ab56164) at 1/1000 dilution

    Lane 1 : mouse small intestine
    Lane 2 : mouse liver

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : goat anti-rabbit secondary antibody at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Additional bands at: 57 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 40 seconds


    Blocked with 5% milk for 1 hour at 25C

    See Abreview

  • Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded human pancreas sections labelling RIP3 with ab56164 at 3 - 12 µg/mL. The secondary antibody used was a donkey anti-rabbit antibody at 1/200. The exposure time was 0.2 seconds at magnification of 20X. 

  • RIP3 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to RIP3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56164.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 65kDa; RIP3.
  • All lanes : Anti-RIP3 antibody (ab56164) at 1.25 µg/ml

    Lane 1 : Molecular weight markers
    Lane 2 : Jurkat cell lysate at 10 µg

    Secondary
    All lanes : HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 57 kDa
    Observed band size: 57 kDa



    Gel concentration: 12%

References

This product has been referenced in:
  • Tang MB  et al. Anisomycin prevents OGD-induced necroptosis by regulating the E3 ligase CHIP. Sci Rep 8:6379 (2018). Read more (PubMed: 29686306) »
  • Wójcik M  et al. Expression of receptor interacting protein 1 and receptor interacting protein 3 oval cells in a rat model of hepatocarcinogenesis. Exp Ther Med 15:4448-4456 (2018). Read more (PubMed: 29731829) »
See all 23 Publications for this product

Customer reviews and Q&As

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1-10 of 26 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 24 2018

Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 24 2018

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

Abcam user community

Verified customer

Submitted Dec 24 2018

Abreviews
Application
Western blot
Sample
Pig Cell lysate - whole cell (Brain Endothelial Cells)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
30 µg
Specification
Brain Endothelial Cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Sep 17 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
BHK-21 (Kidney Fibroblasts)

Dr. Yi-Chieh Perng

Verified customer

Submitted Apr 11 2017

Application
Western blot
Sample
Chinese Hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MES buffer))
Loading amount
50000 cells
Specification
Chinese hamster ovary (CHO) cells
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 12 2016

Application
Western blot
Sample
Rat Cell lysate - whole cell (H9C2 (myoblasts))
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
H9C2 (myoblasts)

Abcam user community

Verified customer

Submitted Nov 18 2016

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Mouse embryonic fibroblasts (MEFs)

Abcam user community

Verified customer

Submitted Oct 27 2016

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Total protein in input
5e+006 cells
Treatment
100 units/mL human IFNbeta
Immuno-precipitation step
Protein G
Specification
A549 (human lung epithelium cells)

Abcam user community

Verified customer

Submitted Oct 27 2016

Application
Immunoprecipitation
Sample
Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Total protein in input
5e+006 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Canine Kidney Epithelial MDCK cells

Abcam user community

Verified customer

Submitted Oct 25 2016

1-10 of 26 Abreviews

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

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