Ubiquitin Antibody Review

Introduction

a 76-amino acid protein, is covalently attached to abnormal and short-lived proteins, thus marking them for ATP-dependent proteolysis in eukaryotic cells. PMID: 8430776
involved in intracellular protein metabolism, but may also have extracellular roles in host defense and immunomodulation. PMID: 16926027
involved in neurodegeneration and various stress responses in the brain. PMID: 15847821
the founding member of a family of structurally conserved proteins that regulate a host of processes in eukaryotic cells. PMID: 15571809
a highly conserved protein essential to an important pathway that targets proteins for proteolysis. Ubiquitin is an immunophilin. PMID: 11805719
Ubiquitination is a universal protein degradation pathway in which the molecules of 8.5-kDa proteolytic peptide ubiquitin are covalently attached to the epsilon-amino group of the substrate's lysine residues. PMID: 11945225

Normal Expression

ubiquitin is highly concentrated in the post-synaptic domain of human neuromuscular junctions. PMID: 1321352
Immunohistochemical localization revealed that differences in ubiquitin pools derived from the relative content of red (oxidative) vs white (glycolytic) fibers, with the former exhibiting a higher content of ubiquitin conjugates. Subsequent immunogold labeling demonstrated statistically significant enhanced localization of ubiquitin conjugates to the Z-lines in both red and white muscle fiber types. PMID: 2835410
Ubiquitin is found within the cytoplasm, nucleus, microvilli, autophagic vacuoles, and lysosomes. PMID: 8430776
Ubiquitin was identified. It was expressed at high levels in old fast-twitch muscles but at low levels in young fast-twitch muscles. PMID: 15081892
Ubiquitin and UbcH7 were observed in both nucleus and cytoplasm of normal, "premalignant" and malignant tissues of oral mucosa using immunoelectron microscopy. However there was no change in their distribution pattern. PMID: 11754502
Ubiquitin was detected within both the cytoplasm and nucleus of the cytotrophoblast layer only. Both monomeric and conjugated forms of ubiquitin were detected. The relative abundance of ubiquitin did not change through gestation or in the two disorders of pregnancy studied. This is the first report to demonstrate the cell-specific localisation of ubiquitin and ubiquitin-protein conjugates in the human cytotrophoblast and provides supportive evidence that ubiquitin may be important during placental development. PMID: 11742419

Abnormal Expression

Using an immunocytochemical method to localise antibodies to ubiquitin, filamentous inclusion bodies were seen in spinal anterior horn neurones in cases of motor neurone disease (MND) but not in any control cases. PMID: 2853853
amyotrophic lateral sclerosis (ALS) neurons exhibited ubiquitin positive inclusions and areas of strong immunoreaction for proteasome. PMID: 16806703
ubiquitin is increased in serum after trauma and might regulate immune functions. PMID: 16003067
an increase in the ubiquitin (Ub)-immunoreactive structures in the midbrain in acute deaths from asphyxiation and in fires in adult subjects. PMID: 15847823
Ub-immunoreactivity was clearly observed in the nuclei of the PGM neurons, showing no postmortem interference or age-dependency. A higher value was observed in blunt injuries, fire fatalities and also in saltwater drowning, hyperthermia and delayed head injury deaths. PMID: 15847822
Using unilaterally 6-hydroxydopamine-denervated rats and mass spectrometry profiling directly on brain tissue sections, we here report for the first time an increased level of unconjugated ubiquitin specifically in the dorsal striatum of the dopamine depleted hemisphere. PMID: 15822896
Both the ubiquitin protein and apoptosis were expressed in both glandular and stromal cells throughout the menstrual cycle of the eutopic endometrium, in which ubiquitin exhibited a cyclic expression, reaching a peak in late proliferative phase. In contrast, ubiquitin was predominantly expressed in cells of stromal origin in endometriosis, was no longer regulated by a cyclic pattern and was associated with an aberrant level of cell survival. PMID: 15447789
the levels of free ubiquitin (FUb) and multi-ubiquitin chain (MUC) were up-regulated in colorectal cancer and were correlated with the pathological findings. PMID: 15194223
Nine cases of atypical Pick's disease without Pick bodies were investigated immunohistochemically. Ubiquitin (ub)-positive and tau-negative structures were mainly found in the cerebral cortex and hippocampal dentate gyrus. PMID: 15068171
Sections of brains from control mice showed virtually no immunostaining for ubiquitin, whereas sections from SOD1G93A transgenic mice contained numerous granular or linear deposits of ubiquitin. PMID: 12870267
Ubiquitin-positive inclusions (UbIs) were located in the cytoplasm of the ependymal cells, and were round to oval in shape, measuring 4-11 microm in diameter. PMID: 12750929
All cases with dementia had numerous ubiquitin-positive inclusions in dentate granule cells, whereas involvement of the neocortex was more variable. PMID: 12536219
Immunohistochemically, intraneuronal ubiquitin-positive, tau and alpha-synuclein -negative inclusions were present in the small neurons of the dentate gyrus, frontal cortices and putamen. PMID: 12471466
Ubiquitin (Ub) was positive in nuclei and cytoplasm of hepatocellular carcinoma (HCC) in immunohistochemistry. The labeling index (L.I.) of ubiquitination was significantly higher with HCC than with other chronic liver diseases and tended to correlate with the lack of poorly-differentiated of HCC. PMID: 12458564
In murine skin wound specimens, neutrophils, macrophages and fibroblasts showed intensive Ub-positive reactions in the nuclei. In the human wound specimens with wound ages between 4 h and 1 day, neutrophils with strong intranuclear positive reactions for Ub were observed. With increasing wound ages, the nuclei of macrophages and fibroblasts were more intensively stained with anti-Ub antibody. PMID: 12376835
The level of Ub immunoreactivity was measured in nucleus and cytoplasm by electron microscopic immunocytochemical and morphometrical methods. We found a significant increase of Ub level in the nucleus and the cytoplasmic area in premalignant atypical acinar cell nodule (AACN) cells and in malignant adenocarcinoma in situ (CIS) cells at month 20 after initiation. PMID: 11693988
Ub-IR cytoplasmic and nuclear staining was significantly greater in both motor neurones and interneurones of MND patients than controls. Ub-IR was less frequent in motor neurone disease (MND) cases in which a great loss of neurones was observed. Ub-IR was significantly more frequent in motor neurones than interneurones, both in patients and controls. Ub-IR inclusions and skeins were only observed in motor neurones from MND patients. PMID: 11679087
Ubiquitin was clearly demonstrated in the nuclei of pigmented substantia nigra neurons, showing two intranuclear staining patterns: a type of inclusion (possibly Marinesco bodies) and a diffuse staining. The diffuse staining was significantly more frequently observed in cases of drowning. PMID: 11599772

Function

an immunohistochemical marker of Mallory bodies and alcoholic liver disease. PMID: 1652996
hyperactivity of the ubiquitin-proteasome pathway is involved in the aging process of fast-twitch muscles. In addition, ubiquitin-dependent growth suppression in satellite cells may be associated with the poor healing potential of old skeletal muscles. PMID: 15081892
From the viewpoint of a forensic pathology application, the present study showed that Ub is suitable as a marker of wound age determination. PMID: 12376835
Ubiquitin immunohistochemistry is a reliable, reproducible, and readily available diagnostic aid for distinguishing Alzheimer's disease from other causes of dementia. PMID: 10786809

Review Articles

Ubiquitin-binding domains. PMID: 17034365
Ubiquitin and ubiquitin-like proteins in cancer pathogenesis. PMID: 16990855
The ubiquitin-proteasome pathway in viral infections. PMID: 16845885
Drug discovery in the ubiquitin-proteasome system. PMID: 16816840
The nuclear ubiquitin-proteasome system. PMID: 16687735
The ubiquitin system and morphogenesis of fungal pathogens. PMID: 16671358
Ubiquitin and SUMO systems in the regulation of mitotic checkpoints. PMID: 16647857
The ubiquitin-proteasome system. PMID: 16595883
Targeting the ubiquitin-proteasome pathway in breast cancer therapy. PMID: 16556079
The ubiquitin-proteasome system in cardiac physiology and pathology. PMID: 16501026
The ubiquitin-proteasome system: focus on the heart. PMID: 16497285
Synapse formation and plasticity: recent insights from the perspective of the ubiquitin proteasome system. PMID: 16427269
The ubiquitin-proteasome system in Huntington's disease. PMID: 16282599
The ubiquitin-proteasome system and cancer. PMID: 16250906
The ubiquitin-proteasome system and skeletal muscle wasting. PMID: 16250905
The ubiquitin-proteasome system and neurodegenerative disorders. PMID: 16250904
Targeting of host-cell ubiquitin pathways by viruses. PMID: 16250903
Transgenic manipulation of the ubiquitin-proteasome system. PMID: 16250902
Fluorescent reporters for the ubiquitin-proteasome system. PMID: 16250901
Response of the ubiquitin-proteasome pathway to changes in muscle activity. PMID: 15886351
Proteomic insights into ubiquitin and ubiquitin-like proteins. PMID: 15701456
Ubiquitin, proteasome and parkin. PMID: 15571819
Targeting the ubiquitin-proteasome pathway in breast cancer. PMID: 15245620, PMID: 15021895
Mechanism of neurodegenerative disease: role of the ubiquitin proteasome system. PMID: 15224658
Motor neuron disease group accompanied by inclusions of unidentified protein signaled by ubiquitin. PMID: 15139588
ubiquitin-proteasome pathway is a new partner for the control of insulin signaling. PMID: 15075914
From neurodegeneration to neurohomeostasis: the role of ubiquitin. PMID: 12792671
The ubiquitin superfamily: members, features, and phylogenies. PMID: 12645912

Applications

Electron Microscopy (IEM)

Several neurodegenerative diseases, including motor neuron disease (MND), are characterized by formation of abnormal cytoskeleton-derived inclusions which contain ubiquitin (Ubq). We have studied the distribution of Ubq in 26 cases of MND with light and electron microscopic immunocytochemistry. PMID: 1648124

ELISA

Ubiquitin in cerebrospinal fluid: a rapid competitive enzyme-linked immunoflow assay. PMID: 7948853
Daily aliquots were drawn from pRBCs (n = 3) for 42 days and plasma was isolated. Ub was measured by ELISA. PMID: 16926027
Ubiquitin immunoreactivity was analyzed by enzyme-linked immunosorbent assay and immunoblotting. PMID: 16003067
Serum ubiquitin levels were studied by ELISA method. RESULTS: The mean serum ubiquitin level (14.13 +/- 1.46 micrograms/mL) in patients with nonalcoholic steatohepatitis was significantly elevated compared to that of the control group (7.66 +/- 0.40 micrograms/mL) (p < 0.001). PMID: 12828075
Serum ubiquitin values were measured by sandwich enzyme-linked immunosorbent assay. PMID: 11194212

Immunocytochemistry (ICC)

distribution of ubiquitin was studied by immunocytochemistry in eight cases of human spongiform encephalopathy and compared with the findings in seven age- and sex-matched cases of Alzheimer's disease and six non-demented control cases. PMID: 8100355
Quantitative studies show that the novel neuropathological technique of anti-ubiquitin immunocytochemistry is more than twice as sensitive as conventional haematoxylin and eosin stains in detecting cortical Lewy bodies. PMID: 2540286
Cortical ubiquitin-positive inclusions in frontotemporal dementia without motor neuron disease: a quantitative immunocytochemical study. PMID: 15170577
We used immunocytochemistry to identify ubiquitin and beta-amyloid-protein in muscle biopsies from patients with three neuromuscular disorders characterized by the presence of rimmed vacuoles in muscle fibres: inclusion body myositis (IBM), familial IBM-like disorder and oculopharyngeal muscular dystrophy (OPMD). PMID: 8268725
Histological sections of cerebral motor cortex, brainstem, and spinal cord from 10 cases of clinically diagnosed motor neurone disease (MND) and 10 control cases were examined by conventional histology and immunocytochemical methods to localise ubiquitin. PMID: 2484732
Overexpression of HDJ-2 protects astrocytes from ischemia-like injury and reduces redistribution of ubiquitin staining in vitro. PMID: 14526221

Immunofluorescence (IF)

We find by immunofluorescence that ubiquitin is present within nucleoli and also demonstrate by immunoprecipitation that complexes associated with pre-rRNA processing factors are ubiquitinated. PMID: 16782897

Immunohistochemistry (IHC)

Pathological heterogeneity of frontotemporal lobar degeneration with ubiquitin-positive inclusions delineated by ubiquitin immunohistochemistry and novel monoclonal antibodies. PMID: 17003490
The optimal methods for carrying out immunohistochemical analyses of paraffin-embedded tissues are described, and examples of all the types of intracellular inclusions detected by ubiquitin immunohistochemistry in the diseases are illustrated. PMID: 16338351
present report describes the neuropathology of 52 brains with Frontotemporal lobar degeneration (FTLD) without tau pathology compared with 10 brains of amyotrophic lateral sclerosis (ALS) without dementia using ubiquitin immunohistochemistry. PMID: 16317257
We performed ubiquitin immunohistochemistry on postmortem tissue, representing a wide range of neuroanatomic structures, in cases of classic motor neuron disease (MND) (n = 20), MND-d (n = 15), and FTD-MND type (n = 15). PMID: 16106222
Using ubiquitin immunohistochemistry and impregnative Nauta method we demonstrated that ubiquitin positivity and Nauta positivity in the neurons affected with ischemic injury in the lumbosacral spinal cord of rabbits and dogs may be of the same origin. PMID: 15568742
Frontotemporal lobar degeneration and ubiquitin immunohistochemistry. PMID: 15305982
Various ubiquitin-positive structures in the hippocampus in diffuse Lewy body disease (n=12) and non-demented aged subjects (n=3) were investigated immunohistochemically. PMID: 9171325
Immunohistochemical detection of ubiquitin-positive intracytoplasmic eosinophilic inclusion bodies in diffuse alveolar damage. PMID: 16722934
we performed immunohistochemical techniques to investigate the changes in ubiquitin expression in the central nervous system of the transgenic mice expressing a human superoxide dismutase 1 mutation (SOD1)G93A. PMID: 12870267
This study provides an immunohistochemical demonstration of the involvement of the ubiquitin- and proteasome-dependent pathway during differentiation and organogenesis in plants. PMID: 11506355
Ubiquitin-immunohistochemical investigation of atypical Pick's disease without Pick bodies. PMID: 9741407
Using immunohistochemistry, we visualized the localization of ubiquitin in the gerbil hippocampus following 3 min of ischemia with or without pretreatment with 2 min of sublethal ischemia and 3 days of reperfusion. PMID: 8397055
we examined the expression of ubiquitin in 14 cases of typical inclusion body myositis, in skeletal muscle specimens from four cases of typical amyotrophic lateral sclerosis, and in muscle specimens from three normal controls. PMID: 8393651
We have investigated the extent and pattern of immunostaining for ubiquitin protein (UBQ) in 60 patients with frontotemporal lobar degeneration (FTLD) with ubiquitin-positive, tau-negative inclusions (FTLD-U). PMID: 17021754
We investigated the immunohistochemical alterations of BDNF, NGF, HSP 70 and ubiquitin in the hippocampus 1 h to 14 days after transient cerebral ischemia in gerbils. PMID: 16810563
Serial sections of formalin-fixed, paraffin-embedded muscle biopsy specimens from 28 Quarter Horse, Paint, and draft-related breeds, aged 0.5-23 years, were treated with periodic acid-Schiff (PAS) stain for glycogen and were immunostained to detect ubiquitin expression. PMID: 16672573
Ubiquitin immunohistochemistry was used for investigation of time dependent changes of ubiquitin in the nerve cells reacting to ischemic/reperfusion damage. PMID: 16670948
Nine cases of atypical Pick's disease without Pick bodies (aPiD) and seven cases of dementia with motor neuron diseases (D-MND) were compared using immunohistochemistry of ubiquitin (ub) and ub-related proteins. PMID: 15641590
Frontotemporal lobar degeneration and ubiquitin immunohistochemistry. PMID: 15305982
Routine and immunohistochemical studies were carried out in a consecutive series of 29 cases of dementia lacking distinctive histology (DLDH). 83% of the cases had ubiquitin-positive inclusions (UPI). PMID: 15178950
Lesions of both infarct-related and noninfarct-related segments from the same patients were examined for the expression and localization of ubiquitin by use of immunohistochemistry and a semiquantitative grading scale. PMID: 12475450

Immunoprecipitation (IP)

We find by immunofluorescence that ubiquitin is present within nucleoli and also demonstrate by immunoprecipitation that complexes associated with pre-rRNA processing factors are ubiquitinated. PMID: 16782897

Western Blotting (WB)

distribution of protein gene product 9.5 (PGP) and ubiquitin immunoreactivities in the ductuli efferentes, ductus epididymidis, and ductus deferens of humans was studied by Western blot analyses and light and electron microscopic immunocytochemistry. PMID: 8828831
Efficient antibodies recognizing ubiquitinated proteins are now available, making ubiquitin immunoblotting a practical tool for research into the many and varied aspects of this extremely interesting posttranslational protein modification. PMID: 15917635
Immunoblot analysis also revealed a decrease in the amount of ubiquitin in the hippocampus ipsilateral to the lesion 1 and 3 days post-lesion. PMID: 11935260
In this investigation we employed immunohistochemistry and immunoblotting techniques to study the distribution and abundance of ubiquitin and ubiquitin-protein conjugates within human placental specimens from throughout gestation. PMID: 11742419