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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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RecombinantRabMAb

Recombinant Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

  • Datasheet
  • SDS
Reviews (9) Submit a question References (3)

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Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Flow Cytometry - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4447] to STUB1/CHIP
  • Suitable for: WB, IP, Flow Cyt, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human

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Protein
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Recombinant Human STUB1/CHIP protein (His tag) (ab206161)
Primary
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Alexa Fluor® 594 Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab206360)

View more associated products

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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • 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.  

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4447
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Ubiquitin ligases
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • E3 Ubiquitin Ligases

Associated products

  • Alternative Versions

    • Anti-STUB1/CHIP antibody [EPR4447] - BSA and Azide free (ab238966)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human STUB1/CHIP protein (His tag) (ab206161)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Mouse
Rat
Human
Application Abreviews Notes
WB (5)
1/10000 - 1/50000. Predicted molecular weight: 35 kDa.
IP (2)
1/120.

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

Flow Cyt
Use at an assay dependent concentration.
ICC/IF (2)
1/250 - 1/500.
IHC-P
1/100.
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.
IHC-P
1/100.

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.
  • Target information above from: UniProt accession Q9UNE7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10273 Human
    • Entrez Gene: 56424 Mouse
    • Entrez Gene: 287155 Rat
    • Omim: 607207 Human
    • SwissProt: Q9UNE7 Human
    • SwissProt: Q9WUD1 Mouse
    • Unigene: 592081 Human
    • Unigene: 491120 Mouse
    • Unigene: 6504 Rat
    see all
  • 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

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

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) 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

  • Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

    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.

  • Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

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

  • Flow Cytometry - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Flow Cytometry - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

    Flow Cytometry analysis of SH-SY5Y (Human epithelial cell line from embryonic kidney) 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.

  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    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

  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)This image is courtesy of an anonymous Abreview
    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 MEF (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 4 : STUB1 knockout MEF (Mouse embryonic fibroblast cell line) 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

  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    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

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

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : ECV-304 (Human urinary bladder cancer cell line) 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



    ECV-304 (derived from T24, discontinued by ATCC)

  • Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
    Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (3)

Publishing research using ab134064? Please let us know so that we can cite the reference in this datasheet.

ab134064 has been referenced in 3 publications.

  • Dema A  et al. Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP. Cells 9:N/A (2020). PubMed: 32164329
  • Pan C  et al. Hsp90B enhances MAST1-mediated cisplatin resistance by protecting MAST1 from proteosomal degradation. J Clin Invest 129:4110-4123 (2019). PubMed: 31449053
  • Li H  et al. WWP2 is a physiological ubiquitin ligase for phosphatase and tensin homolog (PTEN) in mice. J Biol Chem 293:8886-8899 (2018). PubMed: 29685889

Customer reviews and Q&As

Show All Reviews Q&A
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1-9 of 9 Abreviews or Q&A

Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

Average
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Aug 01 2019

Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Aug 01 2019

Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Aug 01 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-STUB1/CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jun 23 2014

Immunoprecipitation abreview for Anti-STUB1/CHIP antibody [EPR4447]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jun 09 2014

Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jun 09 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-STUB1 / CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Mar 05 2013

Immunoprecipitation abreview for Anti-STUB1/CHIP antibody [EPR4447]

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

Abcam user community

Verified customer

Submitted Mar 05 2013

Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jan 03 2013

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