Ubiquitin Antibody (Ubi-1)

Images

 
Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Ubiquitination of PML in response to As2O3. PML-/- MEFs were transiently transfected with the empty vector (EV) or PMLIII (left panel) or contructs encoding PMLIII, ...read more
Immunocytochemistry/ Immunofluorescence: Ubiquitin Antibody (Ubi-1) [NB300-130] - Study of mutant BRAF protein found within the inclusions. BRAFV600E/ubiquitin double-IF labeling. A nucleus with a large inclusion is ...read more
Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Analysis of HEK293 cell lysates using mouse mAb Ubiquitin, dilution 1:1000 (Green). [1] protein standard (Red), [2] cells maintained in normal medium, [3] cells ...read more
Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - FFPE section of cerebral cortex of an Alzheimer patient processed with Ubiquitin antibody using HRP/DAB (Brown) and also stained with haemotoxylin ...read more
Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - Staining of Ubiquitin in hippocampal tissue from an Alzheimer patient.
Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Molecular Analysis of XopD Ub/Ubl Specificity. S. lycopersicum protein extract blotted for total Ub (Ubiquitin; Novus Biologicals) following a 1 hr room temperature ...read more
Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Western blot of Ubiquitin expression on (Lane 1) poly-ubiquitin (lys63 linked), (Lane 2) pure ubiquitin and crude homogenates of adult rat, (Lane 3) cortex, (Lane ...read more
Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - IHC staining of Ubiquitin in human rectal cancer using DAB with hematoxylin counterstain.

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Pm, ZeSpecies Glossary
Applications WB, IB, ICC/IF, IHC, IHC-Fr, IHC-P
Clone
Ubi-1
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

Order Details

Ubiquitin Antibody (Ubi-1) Summary

Immunogen
Purified bovine erythrocyte ubiquitin cross-linked with glutaraldehyde to KLH
Specificity
Recognizes polyubiquitin chains much more strongly than monoubiquitinated molecules or free ubiquitin. Specifically recognizes ubiquitinated cytoplasmic inclusion bodies.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
RPS27A
Purity
Affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:2000
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:2000
  • Immunohistochemistry 1:2000
  • Immunohistochemistry-Frozen 1:2000
  • Immunohistochemistry-Paraffin 1:2000
Application Notes
This Ubiquitin Antibody (Ubi-1) is useful for Western blot, ELISA, Immunocytochemistry/Immunofluorescence, and Immunohistochemistry on both paraffin-embedded and frozen sections. In Western blot a band can be seen at 8.5 kDa. This antibody is excellent for the detection of ubiquitinated inclusions seen in human neurodegenerative diseases such as the neurofibrillary tangles of Alzheimer's disease. Use in immunoblotting reported in scientific literature (PMID: 31727601).
Theoretical MW
8.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Reviewed Applications
Read 3 Reviews rated 4
using
NB300-130 in the following applications:

Publications
Read Publications using
NB300-130 in the following applications:

  • IB
    1 publication
  • 6 publications
  • IHC
    4 publications
  • 1 publication
  • WB
    20 publications

Reactivity Notes

Plant reactivity reported in scientific literature (PMID: 19147500).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS, 50% glycerol (1:1)
Preservative
5mM Sodium Azide
Purity
Affinity purified

Alternate Names for Ubiquitin Antibody (Ubi-1)

  • RPS27A
  • UBA52
  • UBB ubiquitin B
  • UBB
  • UBC
  • Ubiquitin

Background

Ubiquitin is a highly conserved protein of about 8.5 kDa molecular weight, which has a normal role in the targeting of proteins for proteolytic degradation. To perform this function, the protein to be degraded is first covalently attached to the C-terminus of ubiquitin, and the ubiquitinated complex is then recognized by a complex of degradative enzymes. Interestingly, ubiquitin also becomes covalently bonded to many types of pathological inclusions, which appear to be resistant to normal degradation. Therefore, ubiquitin antibodies are very useful for studies of these inclusions. For example, the neurofibrillary tangles and paired helical filaments diagnostic of Alzheimer's disease, Lewy bodies seen in Parkinson's disease, and Pick bodies found in Pick's disease are all heavily ubiquitinated and can be readily visualized with ubiquitin antibodies.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB300-130
Species: Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Pm, Ze
Applications: WB, IB, ICC/IF, IHC, IHC-Fr, IHC-P

Publications for Ubiquitin Antibody (NB300-130)(32)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Drosophila, Plant.

We have publications tested in 5 applications: IB, ICC/IF, IHC, IHC-Fr, WB.


Filter By Application
IB
(1)
ICC/IF
(6)
IHC
(4)
IHC-Fr
(1)
WB
(20)
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Filter By Species
Human
(15)
Mouse
(5)
Rat
(2)
Drosophila
(2)
Plant
(1)
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Showing Publications 1 - 10 of 32. Show All 32 Publications.
Publications using NB300-130 Applications Species
Schwertheim S, KAlsch J, Jastrow H et al. Characterization of two types of intranuclear hepatocellular inclusions in NAFLD Sci Rep Oct 6 2020 [PMID: 33024131] (ICC/IF, Human) ICC/IF Human
Thurner L, Hartmann S, Bewarder M et al. Identification of the atypically modified autoantigen Ars2 as the target of B-cell receptors from activated B cell-type diffuse large B-cell lymphoma Haematologica Jul 16 2020 [PMID: 32675228] (Human) Human
Chen Z, Wasney G A et al. Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12. Open Biol Jan 6 2020 [PMID: 32574548] (WB, Human) WB Human
Yoon YM, Lee JH, Song KH et al. Melatonin-stimulated exosomes enhance the regenerative potential of chronic kidney disease-derived mesenchymal stem/stromal cells via cellular prion proteins J. Pineal Res. Jan 27 2020 [PMID: 31989677]
Schultz E, Jones T, Clamp H, Barr K New insights into intranuclear inclusions in thyroid carcinoma: Association with autophagy and with BRAFV600E mutation PLoS ONE Dec 16 2019 [PMID: 31841566] (ICC/IF, Human) ICC/IF Human
Sn S, Pandurangi J, Murumalla R et al. Small molecule modulator of aggrephagy regulates neuroinflammation to curb pathogenesis of neurodegeneration EBioMedicine Nov 11 2019 [PMID: 31727601] (IB, Human) IB Human
Swatek KN, Usher JL, Kueck AF et al. Insights into ubiquitin chain architecture using Ub-clipping Nature Aug 1 2019 [PMID: 31413367] (WB, Human) WB Human
Aalto AL, Mohan AK, Schwintzer L et al. M1-linked ubiquitination by LUBEL is required for inflammatory responses to oral infection in Drosophila. Cell Death Differ. Jul 19 2018 [PMID: 30026495] (WB, Drosophila) WB Drosophila
Gladkova C, Maslen SL, Skehel JM, Komander D Mechanism of Parkin activation by PINK1 Nature. 2018 Jun 05 [PMID: 29995846] (WB) WB
Gogoi M, Ravikumar V, Dixit NM, Chakravortty D. Salmonella escapes K63 ubiquitination mediated MHC II endosomal proteolysis via modulation of endosomal acidification in dendritic cells. Pathog Dis 2017 Dec 26 [PMID: 29293966] (WB, ICC/IF) WB, ICC/IF
Show All 32 Publications.

Reviews for Ubiquitin Antibody (NB300-130) (3) 43

Average Rating: 4
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Mouse, Human and Mouse.

Reviews using NB300-130:
Filter by Applications
WB
(2)
IF
(1)
All Applications
Filter by Species
Mouse
(2)
Human and Mouse
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot Ubiquitin NB300-130
Enlarge
5
reviewed by:
Anonymous
WB Mouse 03/27/2018
View

Summary

ApplicationWestern Blot
Sample TestedMouse skeletal muscle homogenate
SpeciesMouse
Lot1027092-3

Comments

CommentsWestern Blot: Ubiquitin Antibody (Ubi-1) [NB300-130SS] - Total protein from Mouse skeletal muscle homogenate, separated on a 4-12% gel by SDS-PAGE, transferred to nitrocellulose membrane and blocked in 5% non-fat milk for 1h at room temperature. The membrane was probed with anti-Myogenin 1:1000 in non-fat milk.
  4
reviewed by:
Rune Damgaard
WB Human and Mouse 08/04/2017
View

Summary

ApplicationWestern Blot
Sample Tested293 whole cell lysate, Hela whole cell lysate,HCT116 whole cell lysate,MEF whole cell lysate,mouse liver lysate,mouse bone marrow-derived macrophage cells
SpeciesHuman and Mouse
Lot1027092-2

Comments

CommentsWorks well for WB on total cell lysates (boiling the membrane (nitrocellulose) can increase signal drastically if it is low, e.g. when blotting on primary cells/tissues), and it works really well for WB on Ubiquitin pulldowns, e.g. after a TUBE (tandem ubiquitin-binding entity) pulldown.
  3
reviewed by:
Anonymous
IF Mouse 05/27/2009
View

Summary

ApplicationImmunofluorescence
Sample Testedprimary mouse neuronal culture
SpeciesMouse
Lot120103

Blocking

Blocking DetailsBlocking Buffer: Fetal Bovie Serun, Blocking Time: 30 minutes, Blocking Temp: 22 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Incubation Dilution Buffer: FBS, Primary Ab Dilution Ratio: 1/1000, Incubation Time: 24hrs, Incubation Temp: 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: Alexa 488

Product General Protocols

View specific protocols for Ubiquitin Antibody (NB300-130): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Ubiquitin Antibody (NB300-130). (Showing 1 - 1 of 1 FAQs).

  1. Regarding the WB image on datasheet of NCX1 antibody (6H2) (NB300-127), can you confirm if lane 2 is a rabbit heart sample? In addition, can you provide the dilution fold and loading mount used to obtain this image? Lastly, would you happen to know if t
    • A protein stabilizer has not been added to the antibody NB300-130. However, note that this antibody is supplied as ascites fluid, not as a purified antibody. If you are not familiar with this format, the following descriptions will be useful: Ascites fluid is a complex solution containing multiple proteins and background antibodies as well as the antibody of interest. Ascites contains the antibody produced by the hybridoma cells as well as many other proteins, including background antibodies produced by the mice and albumin. Antibody concentrations in ascites are typically high, but an accurate concentration of the target antibody cannot be readily determined. Therefore, unfortunately you cannot determine the concentration of the ubiquitin antibody within NB300-130 as sold. This is why this product's pack sizes are volumes rather than masses, and why we state on our datasheet: 'This product is unpurified. The exact concentration of antibody is not quantifiable.' All I can suggest is that you carry out antigen affinity chromatography of the antibody, and then determine its concentration.

Secondary Antibodies

 

Isotype Controls

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Blogs on Ubiquitin.

There's an autophagy for that!
By Christina Towers, PhDA critical mechanism that cells use to generate nutrients and fuel metabolism is through a process called autophagy.  This process is complex and involves over 20 different proteins, most of which are highly conserved acro...  Read full blog post.

Article Review: Glucose-induced transcriptional regulation in cancer
Epigenetic mechanisms have been implicated in many physiological and pathophysiological processes. Among these, histone modifications including methylation, phosphorylation, acetylation and ubiquitination, significantly modify gene expression. In c...  Read full blog post.

PINK1: All work and no fun
The protein PINK1 is a mitochondrial-located serine/threonine kinase (PTK) that maintains organelle function and integrity. It not only protects organelles from cellular stress, but it also uses the selective auto-phagocytosis process for cleaning and...  Read full blog post.

Ubiquitin-Mediated Degradation of Cellular Proteins: The Kiss of Death
Ubiquitin is an abundant and essential cellular 9-kd protein that is conserved across evolution from yeast to humans. Ubiquitin is used by cells as a covalent modifier of other proteins both to activate their function and to target them for degradatio...  Read full blog post.

Using Ubiquitin Antibodies in Various Disease Research
Ubiquitin is a small, highly conserved protein which plays an important role in protein breakdown, covalently bonding to proteins to mark them for proteolytic degradation in a process called ubiquitination. Ubiquitin also binds to inclusion bodies (ac...  Read full blog post.

The Heat is On: Heat Shock Proteins and the Link to Cancer
Novus Biologicals offers an extensive antibody catalog targeting heat shock proteins (HSPs). A large protein group covering a number of families, the HSPs are functionally related by their dramatic upregulation in response to stress. Stress triggers m...  Read full blog post.

The Latest Research on IBR-type E3 Ubiquitin Ligases
E3 ubiquitin ligases are standards in most antibody catalogs. These proteins are essential to the process of ubiquitination, which is expressed in protein pathways throughout the body and is often linked to disease states. It is widely used as a bioma...  Read full blog post.

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Recent Reviews

4
5
1
4
1
3
1
2
0
1
0

Anonymous
03/27/2018
Application: WB
Species: Mouse

 
Rune Damgaard
08/04/2017
Application: WB
Species: Human and Mouse

 
Anonymous
05/27/2009
Application: IF
Species: Mouse

Bioinformatics

Gene Symbol RPS27A
Entrez
Uniprot