Storage Buffer | PBS pH7.4, 50% glycerol, 0.09% sodium azide *Storage buffer may change when conjugated |
Storage Temperature | -20ºC, Conjugated antibodies should be stored according to the product label |
Shipping Temperature | Blue Ice or 4ºC |
Purification | Peptide Affinity Purified |
Clonality | Polyclonal |
Specificity | Detects ~10kDa. It also recognizes ubiquinated proteins. |
Cite This Product | StressMarq Biosciences Cat# SPC-119, RRID: AB_2285248 |
Certificate of Analysis | A 1:1000 dilution of SPC-119 was sufficient for detection of free ubiquitin in 15 µg of HeLa lysate by ECL immunoblot analysis using Donkey anti-rabbit IgG:HRP as the secondary antibody. |
Alternative Names | Polyubiquitin B Antibody, RPS27A Antibody, UBA52 Antibody, UBB Antibody, UBC Antibody, ubiquitin B Antibody |
Research Areas | Alzheimer's Disease, Cell Signaling, Neurodegeneration, Neuroscience, Post-translational Modifications, Ubiquitination |
Cellular Localization | Cell membrane, Cytoplasm, Nucleus |
Accession Number | NP_776558.1 |
Gene ID | 281370 |
Swiss Prot | P0CG53 |
Scientific Background | 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 8.5 kDa. 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(3, 4). Ubiquitination also participates in the internalization and degradation of plasma membrane proteins such as some of the TCR subunits while still ER-membrane associated (5). Ubiquitin also plays a role in regulating signal transduction cascades through the elimination inhibitory proteins, such as IκBα and p27 (6). |
References |
1. Wilkinson K.D. (1995) Annu. Rev. Nutr. 15:161-189. 2. Smalle J., Vierstra R.D. (2004) Anu Rev Plant Biol. 55: 555-590. 3. Bonifacino J.S., et al. (1998) Annu Rev Cell Dev Biol. 14: 19-57. 4. Boston Biochem: "Ubiquitin Proteasome Pathway Overview” http://www.bostonbiochem.com/upp.php 5. Yang M., et al. (1998) J Exp Med. 187: 1835-1846. 6. Chen Z.J., et al. (1996) Cell 84: 853-862. |
Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: Heat Shocked Cervical cancer cell line (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite. Heat Shocked at 42°C for 1h.
Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative Solution. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Cytoplasm.
Immunohistochemistry analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:100000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT. This image was produced using an amplifying IHC wash buffer. The antibody has therefore been diluted more than is recommended for other applications..
Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119). Tissue: Cervical cancer cell line (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Ubiquitin Polyclonal Antibody (SPC-119) at 1:200 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Ubiquitin Antibody. (C) Composite.
StressMarq Biosciences :
Based on validation through cited publications.