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Rabbit Recombinant Monoclonal antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 129 publications.


Images

Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (AB140601), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (AB140601), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (AB140601), expandable thumbnail
  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (AB140601), expandable thumbnail
  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (AB140601), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Expected
Expected
Rat
Tested
Expected
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/200 - 1/10000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

1/200 - 1/10000

Notes

-

Species

Rat

Dilution info

1/200 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/20 - 1/1000

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/250

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

1 products for Alternative Version

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Rabbit Recombinant Monoclonal antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 129 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP8589

Purification technique

Affinity purification Protein A

Specificity

This antibody only recognizes polyubiquitin chains formed by Lys-48 (K48) residue linkage. This antibody can detect the target in mouse and rat cell lines and induced tissues.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Ubiquitin is a small regulatory protein found in almost all tissues of eukaryotic organisms. It has a molecular weight of approximately 8.6 kilodaltons. It functions mechanically by attaching to proteins through a process called ubiquitination which involves the formation of an isopeptide bond. Ubiquitin molecules can form polyubiquitin chains through different lysine residues such as K48 and K63 that determine their function. These chains label substrate proteins for various fates including degradation. Ubiquitin is expressed ubiquitously in cells reflecting its essential role in maintaining protein homeostasis.

Biological function summary

The ubiquitin system plays a critical role in regulating protein turnover and quality control within cells. It is part of a larger complex known as the ubiquitin-proteasome system (UPS) which is responsible for degrading proteins that need to be turned over. This process is essential for cell cycle control response to oxidative stress and DNA repair. Ubiquitin's role in tagging proteins for degradation or signaling allows cells to respond quickly to changes in their environment and maintain balance.

Pathways

Ubiquitin functions in several important biological pathways including the Wnt and NF-kB pathways. In the Wnt signaling pathway ubiquitination modulates the stability of key components thereby affecting the pathway's overall activity. In the NF-kB signaling pathway ubiquitin labels inhibitor proteins for degradation which releases and activates NF-kB. These pathways highlight ubiquitin's interaction with proteins such as beta-catenin in Wnt and IkB in NF-kB illustrating how it regulates diverse cellular processes.

Associated diseases and disorders

The dysfunction of the ubiquitin system is linked to neurodegenerative diseases and cancers. Ubiquitin-related defects in protein degradation can lead to the buildup of unwanted proteins contributing to conditions like Parkinson's disease. Connections with cancer are evident as ubiquitin controls cell cycle proteins and aberrant ubiquitination may drive tumor growth and progression. The protein p53 known to be controlled by ubiquitination plays a significant role in cancer related mechanisms when dysregulated. Understanding and targeting ubiquitin-related pathways may provide new therapeutic opportunities for treating these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Blocking and diluting buffer: 5% NFDM/TBST. This image is produced useing purified ab140601.

    All lanes: Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601) at 1/1000 dilution

    All lanes: K48-linked-Ub2-7recombinant protein lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 26 kDa, 35 kDa, 38 kDa, 40 kDa, 58 kDa, 79 kDa

    Observed band size: 26 kDa, 38 kDa, 39 kDa, 42 kDa, 78 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Purified ab140601 staining Ubiquitin (linkage-specific K48) in MCF7 (Human breast adenocarcinoma cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a dilution of 1/1000. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 was used as a counterstain for primary antibody Anti-67kDa Laminin Receptor antibody [EPR8469] ab133645 at 1/2000. DAPI was used as a nuclear counterstain and PBS as a negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Purified ab140601 staining Ubiquitin (linkage-specific K48) in human endometrium carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Blocking and diluting buffer: 5% NFDM/TBST. This image is produced useing purified ab140601.

    All lanes: Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601) at 200 µg

    Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Lane 2: 293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 3: Mouse heart lysate at 20 µg

    Lane 4: Rat heart lysate at 20 µg

    Lane 5: C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

    Lane 6: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 26 kDa

  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Observed band is above 60kDa

    All lanes: Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601) at 1/200 dilution

    Lane 1: PC-12 at 20 µg

    Lane 2: C6 at 20 µg

    Lane 3: L6 at 20 µg

    Lane 4: C2C12 at 20 µg

    Lane 5: Neuro-2a at 20 µg

    Lane 6: NIH3T3 at 20 µg

    Lane 7: SP2/0 at 20 µg

    Lane 8: Raw264.7 at 20 µg

    Lane 9: B16-F0 at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 26 kDa

  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Observed band is above 60kDa

    All lanes: Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601) at 1/200 dilution

    Lane 1: HEK293 at 10 µg

    Lane 2: Jurkat at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 26 kDa

  • Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    All lanes: Western blot - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601) at 1/1000 dilution

    Lane 1: K6-linked-Ub2 recombinant protein at 0.01 µg

    Lane 2: K27-linked-Ub2 recombinant protein at 0.01 µg

    Lane 3: K29-linked-Ub2 recombinant protein at 0.01 µg

    Lane 4: K11-linked-Ub2 recombinant protein at 0.01 µg

    Lane 5: K48-linked-Ub2-7recombinant protein at 0.01 µg

    Lane 6: K63-linked-Ub2-7 recombinant protein at 0.01 µg

    Lane 7: K33-linked-Ub2 recombinant protein at 0.01 µg

    Lane 8: monoubiquitin recombinant protein at 0.01 µg

    Secondary

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

    Predicted band size: 26 kDa

    Observed band size: 17-60 kDa

  • Flow Cytometry (Intracellular) - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Overlay histogram showing HeLa cells stained with ab140601 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab140601, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    ICC/IF image of ab140601 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab140601 at 10μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) 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.

  • Immunohistochemistry - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601), expandable thumbnail

    Immunohistochemistry - Anti-Ubiquitin (linkage-specific K48) antibody [EP8589] (ab140601)

    Immunohistochemical analysis of formalin fixed paraffin embedded human endometrial carcinoma labelling Ubiquitin (linkage-specific K48) with ab140601 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32 mins.

    ab140601 anti-Ubiquitin (linkage-specific K48) [EP8589] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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