Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab80129 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 Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 193 kDa (predicted molecular weight: 193 kDa).

Target

  • Function
    E3 ubiquitin-protein ligase involved in DNA repair. Upon genotoxic stress, accepts ubiquitin from the UBE2N-UBE2V2 E2 complex and transfers it to 'Lys-164' of PCNA which had been monoubiquitinated by UBE2A/B-RAD18, promoting the formation of non-canonical poly-ubiquitin chains linked through 'Lys-63'.
  • Tissue specificity
    Broadly expressed.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Belongs to the SNF2/RAD54 helicase family.
    Contains 1 H15 (linker histone H1/H5 globular) domain.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 1 PHD-type zinc finger.
    Contains 1 RING-type zinc finger.
  • Domain
    The RING finger mediates E3 ubiquitin ligase activity.
  • Information by UniProt
  • Database links
  • Alternative names
    • bA545I5.2 antibody
    • E3 ubiquitin-protein ligase SHPRH antibody
    • FLJ27258 antibody
    • FLJ37625 antibody
    • FLJ45012 antibody
    • FLJ90837 antibody
    • histone-linker antibody
    • KIAA2023 antibody
    • MGC134886 antibody
    • PHD and RING finger domain-containing helicase antibody
    • Shprh antibody
    • SHPRH_HUMAN antibody
    • SNF2 antibody
    • SNF2 histone linker PHD RING helicase antibody
    see all

Images

  • Anti-SHPRH antibody (ab80129) at 1/1000 dilution + HEK293 whole cell lysate at 50 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 193 kDa
    Observed band size : 175 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ICC/IF image of ab80129 stained SHSY5Y 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 (ab80129, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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 100% methanol fixed (5 min) SHSY5Y cells at 5µg/ml.
  • Anti-SHPRH antibody (ab80129) at 1 µg/ml + SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 193 kDa
    Observed band size : 104,193 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 172 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutesThis antibody was raised against an immunogen that is predicted to recognize isoforms (1,2,3,4 and 5) of E3 ubiquitin-protein ligase SHPRH. The predicted molecular weights of isoforms (1,2,3,4 and 5) are 193 kDa, 104kDa, 17 kDa, 190 kDa and 122 kDa respectively.

References

This product has been referenced in:
  • Qu Y  et al. Axitinib blocks Wnt/ß-catenin signaling and directs asymmetric cell division in cancer. Proc Natl Acad Sci U S A 113:9339-44 (2016). WB . Read more (PubMed: 27482107) »

See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116 cell lines)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
HCT116 cell lines
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Jul 14 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 30 2014

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

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