Overview:
| Product Name | Ubiquitin Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Description | Mouse Anti-Bovine Ubiquitin Monoclonal IgG2a Kappa | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Species Reactivity | Human, Mouse, Rat, Bovine | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Applications | WB, ICC/IF, ELISA | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Antibody Dilution | WB (1:1000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Host Species | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen Species | Bovine | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen | Native bovine ubiquitin, conjugated to KLH | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Concentration | 1 mg/ml | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Conjugates |
Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
StreptavidinProperties:
Streptavidin Datasheet Biotin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| R-PE (R-Phycoerythrin) | ||
Overview:
R-PE Datasheet | ![]() | Optical Properties: λex = 565 nm λem = 575 nm εmax = 2.0×106 Φf = 0.84 Brightness = 1.68 x 103 Laser = 488 to 561 nm Filter set = TRITC |
Properties
| Storage Buffer | PBS pH7.4, 50% glycerol, 0.09% sodium azide |
| Storage Temperature | -20ºC |
| Shipping Temperature | Blue Ice or 4ºC |
| Purification | Protein G Purified |
| Clonality | Monoclonal |
| Clone Number | 6C11-B3 |
| Isotype | IgG2a Kappa |
| Specificity | Detects ~10kDa. |
| Cite This Product | StressMarq Biosciences Cat# SMC-171, RRID: AB_1659518 |
| Certificate of Analysis | 1 µg/ml of SMC-171 was sufficient for detection of ubiquitin in 10 µg of Hela Lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody. |
Biological Description
| Alternative Names | Polyubiquitin B Antibody, RPS27A Antibody, UBA52 Antibody, UBB Antibody, UBC Antibody, ubiquitin B Antibody |
| Research Areas | Cell Signaling, Post-translational Modifications, Ubiquitination |
| Cellular Localization | 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 8.5kDa. 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). |
| References |
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” http://www.bostonbiochem.com/upp.php 4. Yang M., et al. (1998) J Exp Med. 187: 1835-1846. 5. Chen Z.J., et al. (1996) Cell 84: 853-862. |
Product Images
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 6C11-B3 (SMC-171). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-171) at 1:100 for 12 hours at 4°C. Secondary Antibody: APC Goat Anti-Mouse (red) 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.
Western Blot analysis of Human Cell lysates showing detection of Ubiquitin protein using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 6C11-B3 (SMC-171). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-171) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Ubiquitin Monoclonal Antibody, Clone 6C11-B3 (SMC-171). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Ubiquitin Monoclonal Antibody (SMC-171) 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.
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| ATTO 565 | ||
Overview:
ATTO 565 Datasheet | ![]() | Optical Properties: λex = 563 nm λem = 592 nm εmax = 1.2×105 Φf = 0.9 τfl = 3.4 n Brightness = 10 Laser = 532 nm Filter set = TRITC |
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② 应取pH8.0,这样可使核苷酸带较多负电荷,利于吸附于阴离子交换树脂柱。虽然pH 11.4时核苷酸带有更多的负电荷,但pH过高对分离不利。
③ 当不考虑树脂的非极性吸附时,根据核苷酸负电荷的多少来决定洗脱速度,则洗脱顺序为CMP>AMP> GMP > UMP,但实际上核苷酸和聚苯乙烯阴离子交换树脂之间存在着非极性吸附,嘌呤碱基的非极性吸附是嘧啶碱基的3倍。静电吸附与非极性吸附共同作用的结果使洗脱顺序为:CMP> AMP > UMP >GMP。
1. Buffer中离子浓度过大,甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高,也是一个原因















