Overview

  • Product name

    Anti-STUB1/CHIP antibody [EPR4447]
    See all STUB1/CHIP primary antibodies
  • Description

    Rabbit monoclonal [EPR4447] to STUB1/CHIP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human STUB1/CHIP aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • HeLa, MCF-7, 293T and ECV-304 cell lysates; Human skeletal muscle tissue.
  • General notes

    Previously labelled as STUB1.  

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab134064 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 1/10000 - 1/50000. Predicted molecular weight: 35 kDa.
IP 1/120.

For unpurified, use 1/10 - 1/100.

Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.

Target

  • Function

    E3 ubiquitin-protein ligase which targets misfolded chaperone substrates towards proteasomal degradation. Ubiquitinates NOS1 in concert with Hsp70 and Hsp40. Modulates the activity of several chaperone complexes, including Hsp70, Hsc70 and Hsp90. Mediates transfer of non-canonical short ubiquitin chains to HSPA8 that have no effect on HSPA8 degradation. Mediates polyubiquitination of DNA polymerase beta (POLB) at 'Lys-41', 'Lys-61' and 'Lys-81', thereby playing a role in base-excision repair: catalyzes polyubiquitination by amplifying the HUWE1/ARF-BP1-dependent monoubiquitination and leading to POLB-degradation by the proteasome. Mediates polyubiquitination of CYP3A4.
  • Tissue specificity

    Highly expressed in skeletal muscle, heart, pancreas, brain and placenta. Detected in kidney, liver and lung.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Contains 3 TPR repeats.
    Contains 1 U-box domain.
  • Domain

    The TPR domain is essential for ubiquitination mediated by UBE2D1.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
    Auto-ubiquitinated; mediated by UBE2D1 and UBE2D2.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen NY CO 7 antibody
    • Antigen NY-CO-7 antibody
    • C terminus of Hsp70-interacting protein antibody
    • Carboxy terminus of Hsp70 interacting protein antibody
    • Carboxy terminus of Hsp70-interacting protein antibody
    • Carboxy terminus of Hsp70p interacting protein antibody
    • CHIP antibody
    • CHIP_HUMAN antibody
    • CLL associated antigen KW 8 antibody
    • CLL-associated antigen KW-8 antibody
    • E3 ubiquitin protein ligase CHIP antibody
    • E3 ubiquitin-protein ligase CHIP antibody
    • Heat shock protein A binding protein 2 (c terminal) antibody
    • HSPABP2 antibody
    • NY CO 7 antibody
    • PP1131 antibody
    • SDCCAG7 antibody
    • Serologically defined colon cancer antigen 7 antibody
    • STIP1 homology and U Box containing protein 1 antibody
    • STIP1 homology and U box containing protein 1 E3 ubiquitin protein ligase antibody
    • STIP1 homology and U box-containing protein 1 antibody
    • STUB 1 antibody
    • STUB1 antibody
    • UBOX 1 antibody
    • UBOX1 antibody
    see all

Images

  • Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/50000 dilution (purified) + rat brain at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size: 35 kDa
    Observed band size: 35 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of SH-SY5Y cells with purified ab134064 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab134064 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labelling STUB1/CHIP with unpurified ab134064 at 1/100 dilution.

  • Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling STUB1/CHIP with purified ab134064 at 1/120 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (purified) + mouse brain at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size: 35 kDa
    Observed band size: 35 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/20000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 35 kDa
    Observed band size: 35 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • ab134064 (purified) at 1/120 immunoprecipitating STUB1/CHIP in 10 μg HeLa (Lanes 1 and 2, observed at 35 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF-7 cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : ECV-304 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 35 kDa

  • All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/1000 dilution (unpurified)

    Lane 1 : Wild type primary mouse neurons whole cell lysate
    Lane 2 : STUB1 knockout primary mouse neurons whole cell lysate
    Lane 3 : Wild type MEFs whole cell lysate
    Lane 4 : STUB1 knockout MEFs whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 35 kDa


    Exposure time: 25 seconds

    See Abreview

References

ab134064 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Western blot
Sample
Monkey Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 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 Aug 01 2019

Application
Western blot
Sample
Rat Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 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 Aug 01 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 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 Aug 01 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (primary cortical neuron)
Permeabilization
Yes - 0.1% Triton/PBS 5 min
Specification
primary cortical neuron
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 23 2014

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Total protein in input
500 µg
Immuno-precipitation step
Other - Protein G Dynabeads crosslinked to antibody
Specification
mouse embryonic fibroblasts

Abcam user community

Verified customer

Submitted Jun 09 2014

Application
Western blot
Sample
Mouse Cell lysate - whole cell (primary neurons and MEFs)
Gel Running Conditions
Reduced Denaturing (10% Bis-Tris)
Loading amount
20 µg
Specification
primary neurons and MEFs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 09 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
No
Specification
HeLa
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 05 2013

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa)
Total protein in input
1000 µg
Immuno-precipitation step
Protein A/G
Specification
HeLa

Abcam user community

Verified customer

Submitted Mar 05 2013

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

Abcam user community

Verified customer

Submitted Jan 03 2013

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