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Ubiquitin antibody

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Cow (Bovine), Caenorhabditis elegans (C. elegans), Chicken, Fruit Fly (Drosophila melanogaster), Human, Mouse (Murine), Plant
(276), (200), (125), (120), (39), (29), (29), (26), (24), (24), (24), (22), (22), (22), (21), (20), (11), (8), (4), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1)
(221), (171), (6), (5), (3), (1)
Clonality (Clone)
Monoclonal ()
(18), (14), (12), (12), (12), (11), (5), (5), (5), (5), (5), (5), (5), (5), (5), (5), (5), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Western Blotting (WB), ELISA, Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
(307), (196), (138), (87), (86), (73), (47), (33), (24), (12), (11), (9), (4), (3), (2), (1)
Pubmed 14 references available
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Quantity 0.1 mL
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Immunogen Purified Ubiquitin conjugated to KLH with glutaraldehyde.
Clone Ubi-1
Isotype IgG1 kappa
Specificity Recognizes polyubiquitin chains much more strongly than monoubiquitinated molecules or free ubiquitin. Specifically recognizes ubiquitinated cytoplasmic inclusion bodies.
Purification Ascites
Background Ubiquitin is a highly conserved protein of about 8.5 kDa molecular weight, which has anormal role in the targeting of proteins for proteolytic degradation. To perform thisfunction, the protein to be degraded is first covalently attached to the C-terminus ofubiquitin, and the ubiquitinated complex is then recognized by a complex of degradativeenzymes. Interestingly, ubiquitin also becomes covalently bonded to many types ofpathological inclusions, which appear to be resistant to normal degradation. Therefore,ubiquitin antibodies are very useful for studies of these inclusions. For example, theneurofibrillary 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 areall heavily ubiquitinated and can be readily visualized with ubiquitin antibodies. Alternate Names: anti-60S ribosomal protein L40 antibody, anti-HUBCEP52 antibody, anti-RPL40antibody, anti-Ubiquitin 52 amino acid fusion protein antibody, anti-Ubiquitin A52 residueribosomal protein fusion product 1 antibody, anti-Ubiquitin carboxyl extension protein 52antibody, anti-Ubiquitin CEP52 antibody.
Gene Symbol: RPS27A
Gene ID 6233, 22190
Research Area Neurology, Proteolysis / Ubiquitin, Alzheimer's Disease, Proteases, Amino Acids
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.
Recommended dilutions: ELISA 1:100-1:2000, Immunocytochemistry/Immunofluorescence 1:500, Immunohistochemistry 1:500-1:1000, Immunohistochemistry-Frozen 1:500-1:1000, Immunohistochemistry-Paraffin 1:500-1:1000, Western Blot 1:1000-1:5000
Protocol Immunohistochemistry-Paraffin Protocol Specific for NB300-130: Ubiquitin Antibody (Ubi-1)Materials1) 1 Phosphate buffered saline (pH 7.6): NaCl 137mmol/L, KCl 2.7mmol/L, Na2HPO4 4.3mmol/L, KH2PO4 1.4 mmol/L2) Citrate buffer, 0.01 M, pH6.0, Sodium Citrate 3g, Citric acid 0.4g3) 3 % Hydrogen peroxide4) Primary antibody5) Blocking serum (normal serum)6) Biotinylated secondary antibody7) DAB staining kitMethods
1. Dewax and hydration of slides using xylene and EtOH:Dry slides for 20 min in a 60 C ovenAdd Xylene, 2 x 10 min100 %, 95 %, 80 %, and 70 % EtOH, 5 min each EtOH concentration Rinse in PBS, 5'2 Antigen retrieval method (only for paraffin slides)1a. High-pressure antigen retrieval procedure (recommended method)Place slides in a glass slide holder (ensure that the slide holder is completely filled with slides, slides without sections if necessary, to ensure even heating. The entire slide holder is immersed in 1000 mL of Citrate buffer (0.01M, pH6.
0) within a pressure cookerOnce steam is produced, and ONLY when steam is visible, from the pressure cooker (usually 15-20 min), the required high-pressure will have been reached, and slides will be incubated for 2 min.Turn off heat, and allow buffer and slides to cool to room temperatureSlides are then rinsed in PBS for 5 minutes
. Add 3 % hydrogen peroxide solution, 10'at RT, then PBS, 3X5'
. Normal blocking serum, 20'at RT
. Incubate with Primary Ab, 4C overnight or 1.5 hours at 37C
. Rinse with PBS, 3 X 5' each rinse
. Add Biotin-conjugated second antibody, 10'at RT
. Rinse with PBS, 3 X 5' each rinse
. Add Streptavidin-Peroxidase, 10'at RT
. Rinse with PBS, 3 X 5' each rinse
. Staining with DAB solution, 2-5'under microscope
. Stop the reaction by washing in tap water
. Counterstain in Haematoxylin for 3-5 minutes
. 75 %, 80 %, 95 % and 100 % ethanol, 5x2', xylene 2 x 10'
Restrictions For Research Use only
Format Liquid
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
Immunohistochemistry (IHC) image for anti-Ubiquitin antibody (ABIN152476) Immunohistochemical staining of Ubiquitin in hippocampal tissue from an Alzheimer pat...
Western Blotting (WB) image for anti-Ubiquitin antibody (ABIN152476) Staining of ubiquitin using ABIN152476. Lane 1 is mono-ubiquitin and polyubiquitin c...
Product cited in: Stuttmann, Lechner, Guérois et al.: "COP9 signalosome- and 26S proteasome-dependent regulation of SCFTIR1 accumulation in Arabidopsis." in: The Journal of biological chemistry, Vol. 284, Issue 12, pp. 7920-30, 2009 (PubMed).

Dayal, Sparks, Jacob et al.: "Suppression of the deubiquitinating enzyme USP5 causes the accumulation of unanchored polyubiquitin and the activation of p53." in: The Journal of biological chemistry, Vol. 284, Issue 8, pp. 5030-41, 2009 (PubMed).

Watson, Lagow, Xu et al.: "A drosophila model for amyotrophic lateral sclerosis reveals motor neuron damage by human SOD1." in: The Journal of biological chemistry, Vol. 283, Issue 36, pp. 24972-81, 2008 (PubMed).

Cripps, Thomas, Jeng et al.: "Alzheimer disease-specific conformation of hyperphosphorylated paired helical filament-Tau is polyubiquitinated through Lys-48, Lys-11, and Lys-6 ubiquitin conjugation." in: The Journal of biological chemistry, Vol. 281, Issue 16, pp. 10825-38, 2006 (PubMed).

Background publications Lichtenfels, Kellner, Atkins et al.: "Identification of metabolic enzymes in renal cell carcinoma utilizing PROTEOMEX analyses." in: Biochimica et biophysica acta, Vol. 1646, Issue 1-2, pp. 21-31, 2003 (PubMed).

Marti, Wirbelauer, Scheffner et al.: "Interaction between ubiquitin-protein ligase SCFSKP2 and E2F-1 underlies the regulation of E2F-1 degradation." in: Nature cell biology, Vol. 1, Issue 1, pp. 14-9, 1999 (PubMed).

Sternsdorf, Puccetti, Jensen et al.: "PIC-1/SUMO-1-modified PML-retinoic acid receptor alpha mediates arsenic trioxide-induced apoptosis in acute promyelocytic leukemia." in: Molecular and cellular biology, Vol. 19, Issue 7, pp. 5170-8, 1999 (PubMed).

Laroia, Cuesta, Brewer et al.: "Control of mRNA decay by heat shock-ubiquitin-proteasome pathway." in: Science (New York, N.Y.), Vol. 284, Issue 5413, pp. 499-502, 1999 (PubMed).

Verdier, Chruetien, Muller et al.: "Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation. Possible involvement of the ubiquitinated Cis protein." in: The Journal of biological chemistry, Vol. 273, Issue 43, pp. 28185-90, 1998 (PubMed).

Cuervo, Hu, Lim et al.: "IkappaB is a substrate for a selective pathway of lysosomal proteolysis." in: Molecular biology of the cell, Vol. 9, Issue 8, pp. 1995-2010, 1998 (PubMed).

Sternsdorf, Jensen, Will: "Evidence for covalent modification of the nuclear dot-associated proteins PML and Sp100 by PIC1/SUMO-1." in: The Journal of cell biology, Vol. 139, Issue 7, pp. 1621-34, 1998 (PubMed).

Perry, Friedman, Shaw et al.: "Ubiquitin is detected in neurofibrillary tangles and senile plaque neurites of Alzheimer disease brains." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 84, Issue 9, pp. 3033-6, 1987 (PubMed).

Shaw, Chau: "Ubiquitin and microtubule-associated protein tau immunoreactivity each define distinct structures with differing distributions and solubility properties in Alzheimer brain." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, Issue 8, pp. 2854-8, 1988 (PubMed).

Hirano, Kimoto, Shimoyama et al.: "Identification of a neural alpha-catenin as a key regulator of cadherin function and multicellular organization." in: Cell, Vol. 70, Issue 2, pp. 293-301, 1992 (PubMed).

Catalog No. ABIN152476

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