Overview

  • Product name
  • Description
    Rabbit polyclonal to RNF31/HOIP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1050 to the C-terminus of Human RNF31/HOIP.

    Read Abcam's proprietary immunogen policy (Peptide available as ab46321.)

  • Positive control
    • ab46322 gave a positive signal in Jurkat Whole Cell Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab46322 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
ICC/IF
WB
ChIP
  • Application notes
    ChIP: Use at an assay dependent dilution (PMID 19237537.
    ICC/IF: Use at a concentration of 1 µg/ml.
    WB: 1/250. Detects a band of approximately 116 kDa (predicted molecular weight: 119 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      E3 ubiquitin-protein ligase component of the LUBAC complex which conjugates linear polyubiquitin chains in a head-to-tail manner to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation. LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways. Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation. LUBAC is proposed to be recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex. Binds polyubiquitin of different linkage types.
    • Tissue specificity
      Expressed in both normal and transformed breast epithelial cell lines.
    • Sequence similarities
      Contains 2 IBR-type zinc fingers.
      Contains 3 RanBP2-type zinc fingers.
      Contains 1 RING-type zinc finger.
      Contains 1 UBA domain.
    • Domain
      The RanBP2-type zinc fingers mediate the specific interaction with ubiquitin.
    • Cellular localization
      Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • E3 ubiquitin protein ligase RNF31 antibody
      • FLJ10111 antibody
      • FLJ23501 antibody
      • HOIL 1L interacting protein antibody
      • HOIL-1-interacting protein antibody
      • HOIL1L interacting protein antibody
      • HOIP antibody
      • RING finger protein 31 antibody
      • RNF31 antibody
      • RNF31_HUMAN antibody
      • ZIBRA antibody
      • Zinc in between ring finger ubiquitin associated domain antibody
      • Zinc in-between-RING-finger ubiquitin-associated domain protein antibody
      see all

    Images

    • Anti-RNF31/HOIP antibody (ab46322) at 1/250 dilution + Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

      Secondary
      IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 119 kDa
      Observed band size: 116 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 105 kDa (possible isoform), 50 kDa (possible isoform), 60 kDa. We are unsure as to the identity of these extra bands.

    • ICC/IF image of ab46322 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46322, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293 cells at 5µg/ml.
    • Western blot analysis of A549 cell lysate (100μg/lane) RNF31/HOIP with ab46322 at 1/1000. A HRP-conjugated Goat anti-rabbit polyclonal (1/10000) was used as the secondary antibody.

      lane 1-2 : siCT A549
      lane 3-4 : siHOIP A549

      See Abreview

    References

    This product has been referenced in:
    • Dziedzic SA  et al. ABIN-1 regulates RIPK1 activation by linking Met1 ubiquitylation with Lys63 deubiquitylation in TNF-RSC. Nat Cell Biol 20:58-68 (2018). Read more (PubMed: 29203883) »
    • Qiu P  et al. MicroRNA-378 regulates cell proliferation and migration by repressing RNF31 in pituitary adenoma. Oncol Lett 15:789-794 (2018). Read more (PubMed: 29399147) »
    See all 12 Publications for this product

    Customer reviews and Q&As

    1-6 of 6 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A549 Cells)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    100 µg
    Specification
    A549 Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Jan 10 2018

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (primary mouse alveolar type II cells)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    125 µg
    Specification
    primary mouse alveolar type II cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Nov 20 2017

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris, MES buffer)
    Loading amount
    10 µg
    Specification
    Mouse embryonic fibroblasts
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C

    Dr. Paola Marco i Casanova

    Verified customer

    Submitted Mar 28 2014

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293 and hTERT-RPE1)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris, MES buffer)
    Loading amount
    10 µg
    Specification
    HEK293 and hTERT-RPE1
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Mar 28 2014

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK 293)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    30 µg
    Specification
    HEK 293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Aug 12 2011

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

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