Name: Anti-STUB1/CHIP antibody [EPR4447]
Brand: RabMAb
SKU: ab134064
Rating: 4 (9 reviews)

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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.

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: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR4447
Isotype

IgG
Research areas

Alternative Versions
- Anti-STUB1/CHIP antibody [EPR4447] - BSA and Azide free (ab238966)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa whole cell lysate (ab29545)
- 293T whole cell lysate (ab95494)
Recombinant Protein
- Recombinant Human STUB1/CHIP protein (His tag) (ab206161)

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.

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

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
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.
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 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.
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)

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

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
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 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

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

Datasheet

ab134064 has not yet been referenced specifically in any publications.

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

Customer reviews and Q&As

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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

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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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

Applications

Target

Function

Tissue specificity

Pathway

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References

Customer reviews and Q&As

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

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

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

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

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

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

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

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

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

Post-translational
modifications