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Clone
5B9-B3
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Immunogen
Native bovine ubiquitin, conjugated to KLH
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Antibody s full description
Mouse Anti-Bovine Ubiquitin Monoclonal IgG2a Kappa Antibody, Clone: 5B9-B3: Biotin
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Antibody s category
Monoclonal Antibodies
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Antibody s other name
Polyubiquitin B Antibody, RPS27A Antibody, UBA52 Antibody, UBB Antibody, UBC Antibody, ubiquitin B Antibody
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Raised in
Mouse
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Antibody s target
Ubiquitin
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Primary research fields
Cell Signaling, Post-translational Modifications, Ubiquitination
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Brandname
None
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Antibodies applications
WB, ICC/IF, ELISA
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Antibody s reactivity
Human, Mouse, Rat, Bovine
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Antibody s dilutions
WB (1:1000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.
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Purity
Protein G Purified
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Antibody buffer for storage
PBS pH7.4, 50% glycerol, 0.09% sodium azide
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Antibody s concentration
1 mg/ml
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Antibody s specificity
Detects ~10kDa.
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Storage recommendations
-20ºC
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Shipping recommendations
Blue Ice or 4ºC
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Antibody certificate of analysis
1 µg/ml of SMC-160 was sufficient for detection of ubiquitin in 10 µg of Heal Lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.
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Antibody in cell
Cytoplasm , Nucleus
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Tissue specificity
See included datasheet or contact our support service
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Scientific context
Ubiquitin is a small protein that occurs in all eukaryotic cells. The ubiquitin protein itself consists of 76 amino acids and has a molecular mass of about 8.5kDa. Key features include its C-terminal tail and the 7 Lys residues. It is highly conserved among eukaryotic species: Human and yeast ubiquitin share 96% sequence identity (1). The main function of Ubiquitin is to clear abnormal, foreign and improperly folded proteins by targeting them for degradation by the 26S proteosome (2). Ubiquitination represents an essential cellular process affected by a multi-enzyme cascade involving classes of enzymes known as ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s or Ubcs) and ubiquitin-protein ligases (E3s). Ubiquitin is activated in a two-step reaction by an E1 ubiquitin-activating enzyme in a process requiring ATP as an energy source. The initial step involves production of an ubiquitin-adenylate intermediate. The second step transfers ubiquitin to the E1 active site cysteine residue, with release of AMP. This step results in a thioester linkage between the C-terminal carboxyl group of ubiquitin and the E1 cysteine sulfhydryl group. The third step is a transfer of ubiquitin from E1 to the active site cysteine of a ubiquitin-conjugating enzyme E2 via a trans(thio)esterification reaction. And the final step of the ubiquitylation cascade creates an isopeptide bond between a lysine of the target protein and the C-terminal glycine of ubiquitin. In general, this step requires the activity of one of the hundreds of E3 ubiquitin-protein ligases (often termed simply ubiquitin ligase). E3 enzymes function as the substrate recognition modules of the system and are capable of interaction with both E2 and substrate(2, 3). Ubiquitination also participates in the internalization and degradation of plasma membrane proteins such as some of the TCR subunits while still ER-membrane associated (4). Ubiquitin also plays a role in regulating signal transduction cascades through the elimination inhibitory proteins, such as IκBα and p27 (5).
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Bibliography
1. Wilkinson K.D. (1995) Annu. Rev. Nutr. 15:161-189. 2. Bonifacino J.S., et al. (1998) Annu Rev Cell Dev Biol. 14: 19-57. 3. Boston Biochem: "Ubiquitin Proteasome Pathway Overview” more 4. Yang M., et al. (1998) J Exp Med. 187: 1835-1846. 5. Chen Z.J., et al. (1996) Cell 84: 853-862.
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Released date
1-Apr-2009
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Tested applications
To be tested
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Tested reactivity
To be tested
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NCBI number
NP_776558.1
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Gene number
281370
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Protein number
P0CG53
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Antibody s datasheet
Contact our support service
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Representative figure link
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Representative figure legend
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 5B9-B3 (SMC-160). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-160) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Diffuse nuclear and cytoplasmic staining. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite. Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 5B9-B3 (SMC-160). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-160) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Diffuse nuclear and cytoplasmic staining. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite. Western Blot analysis of Human cell lysates showing detection of Ubiquitin protein using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 5B9-B3 (SMC-160). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-160) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Mouse Anti-Ubiquitin Antibody [5B9-B3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HeLa Cells (SMC-160) Mouse Anti-Ubiquitin Antibody [5B9-B3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HeLa Cells (SMC-160) Mouse Anti-Ubiquitin Antibody [5B9-B3] used in Western Blot (WB) on Human cell lysates (SMC-160)
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Warning information
Non-hazardous
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Country of production
Canada
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Total weight kg
1.4
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Net weight g
0.2
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Stock availability
In Stock
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Description
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided. Antibody for research use.
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About
Monoclonals of this antigen are available in different clones. Each murine monoclonal anibody has his own affinity specific for the clone. Mouse monoclonal antibodies are purified protein A or G and can be conjugated to FITC for flow cytometry or FACS and can be of different isotypes.
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Test
Mouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.
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Latin name
Mus musculus
