Name: Anti-Ubiquitin antibody [EPR8830]
SKU: ab134953
Rating: 4 (3 reviews)

Anti-Ubiquitin antibody [EPR8830] is a rabbit recombinant monoclonal antibody that is used to detect Ubiquitin in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Recombinant format for unrivaled batch-batch consistency
- Cited in over 100 publications

Related conjugates and formulations

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Alexa Fluor® 488 Anti-Ubiquitin antibody [EPR8830]
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Alexa Fluor® 647 Anti-Ubiquitin antibody [EPR8830]
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Alexa Fluor® 594 Anti-Ubiquitin antibody [EPR8830]
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Anti-Ubiquitin antibody [EPR8830] - BSA and Azide free
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APC Anti-Ubiquitin antibody [EPR8830]
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Images

Western blot - Anti-Ubiquitin antibody [EPR8830] (AB134953), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
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

	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Expected	Tested
Rat	Tested	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes Monoubiquitin molecular weight Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000 - 1/10000	Notes Monoubiquitin molecular weight Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000 - 1/10000	Notes Monoubiquitin molecular weight Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes For unpurified use at 1/250 - 1/500.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/70	Notes For unpurified use at 1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/800	Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/800	Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/800	Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information UBB methyl

Function

Ubiquitin. Exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in proteotoxic stress response and cell cycle; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
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.
Clone number: EPR8830
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle, Stable for 12 months at -20°C

Notes

Product Specifications
Anti-Ubiquitin antibody [EPR8830] (ab134953) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, WB in human, mouse, rat samples.
Anti-Ubiquitin antibody [EPR8830] (ab134953) specifically detects Ubiquitin (UniProt ID: P0CG47; Molecular weight: 9kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-Ubiquitin antibody [EPR8830] (ab134953) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-Ubiquitin antibody [EPR8830] (ab134953) has been cited over 105 times in peer reviewed journals and is trusted by the scientific community.

Related Products
Antibody clone EPR8830 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, PE, APC, HRP, Alexa Fluor® 568, Alkaline Phosphatase, Alexa Fluor® 594, Alexa Fluor® 555, Alexa Fluor® 750 (ab205467, ab205468, ab303020, ab303021, ab303022, ab307636, ab308798, ab310519, ab312049, ab321094).

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.

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.

16 product images

Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953) at 0.7 µg/mL
Lane 1: Rat brain lysate at 20 µg
Lane 2: Mouse kidney lysate at 20 µg
Lane 3: Rat kidney 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
Observed band size: 8 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunocytochemistry/ Immunofluorescence analysis of JAR (Human placenta choriocarcinoma cell line) cells labeling Ubiquitin with Purified ab134953 at 1:100 dilution (7.2μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling Ubiquitin with Purified ab134953 at 1:800 dilution (0.9 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP)
secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling Ubiquitin with Purified ab134953 at 1:800 dilution (0.9 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP)
secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling Ubiquitin with Purified ab134953 at 1:800 dilution (0.9 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953) at 0.1 µg/mL
Lane 1: JAR (Human placenta choriocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: 293 (Human embryonic kidney epithelial 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
Observed band size: 8 kDa
This image is courtesy of an anonymous customer review.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
ab134953 staining Ubiquitin in Mouse embryo tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer, pH6. Samples were incubated with primary antibody (1/100 in 1% FBS/BSA) for 16 hours at 4°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes .ab134953 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/200 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging using the Licor Odyssey CLx.
All lanes: Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953) at 1/200 dilution
Lane 1: MCF-7 Whole cell lysate at 20 µg
Lane 2: MCF-7 Whole cell lysate + MG-132 (50 uM 90 min) at 20 µg
Lane 3: Mouse Brain at 20 µg
Performed under reducing conditions.
Predicted band size: 26 kDa
Flow Cytometry (Intracellular) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Overlay histogram showing HepG2 cells stained with ab134953 (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 (ab134953, 1/1000 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) (0.1μg/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. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunofluorescent staining of JAR cells labelling Ubiquitin using ab134953 at 1/250 dilution
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Note: Ubiquitin exists as monomeric ubiquitin (8
kDa) or attached to other proteins.
All lanes: Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953) at 1/1000 dilution
Lane 1: 293T cell lysate at 10 µg
Lane 2: HepG2 cell lysate at 10 µg
Lane 3: HeLa cell lysate at 10 µg
Lane 4: JAR cell lysate at 10 µg
Secondary
All lanes: HRP labelled goat anti rabbit at 1/2000 dilution
Predicted band size: 26 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Immunohistochemical analysis of paraffin embedded Human breast carcinoma tissue labelling Ubiquitin with ab134953 at 1/250 dilution
Flow Cytometry (Intracellular) - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Intracellular Flow Cytometry analysis of JAR (Human placenta choriocarcinoma cell line) cells labeling Ubiquitin with purified ab134953 at 1/70 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Ubiquitin antibody [EPR8830] (ab134953) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) (Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) ab320096) at 1/1000 dilution
Lane 1: Untreated PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: PC-3 treated with 30 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 2 hours, whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Lane 4: PC-3 treated with 30 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 2 hours, whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/10000 dilution
Observed band size: 8-200 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Ubiquitin antibody [EPR8830] (ab134953) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) (Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) ab320096) at 1/1000 dilution
Lane 1: Untreated MEF (mouse embryo fibroblast) whole cell lysate (untreated membrane) at 20 µg
Lane 2: MEF treated with 5 μM Valinomycin for 24 hours whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated MEF (mouse embryo fibroblast) whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Lane 4: MEF treated with 5 μM Valinomycin for 24 hours whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/10000 dilution
Observed band size: 8-200 kDa, 36 kDa
Exposure time: 92s
Western blot - Anti-Ubiquitin antibody [EPR8830] (ab134953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Ubiquitin antibody [EPR8830] (ab134953) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) (Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] - Rat IgG2a (Chimeric) ab320096) at 1/1000 dilution
Lane 1: Untreated Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: 293T treated with 20 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 8 hours, whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated PINK1 knockout HEK-293T whole cell lysate (untreated membrane) at 20 µg
Lane 4: PINK1 knockout HEK-293T treated with 20 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 8 hours, whole cell lysate (untreated membrane) at 20 µg
Lane 5: Untreated Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Lane 6: 293T treated with 20 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 8 hours, whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Lane 7: Untreated PINK1 knockout HEK-293T whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Lane 8: PINK1 knockout HEK-293T treated with 20 μM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 8 hours, whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/10000 dilution
Observed band size: 8-200 kDa, 36 kDa
Exposure time: 92s