![StressMarq/Anti-Ubiquitin Antibody [5B9-B3]/SMC-160B-A565/200-µg](images/StressMarq/201710/SMC-160_Ubiquitin_Antibody_5B9-B3_ICC-IF_Human_HeLa-Cells_100x_Composite-150x150.png)
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 BiotinProperties:
Biotin Datasheet HRP (Horseradish peroxidase)Properties:
HRP Datasheet AP (Alkaline Phosphatase)Properties:
AP Datasheet
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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 | 5B9-B3 |
Isotype | IgG2a Kappa |
Specificity | Detects ~10kDa. |
Cite This Product | StressMarq Biosciences Cat# SMC-160, RRID: AB_2285240 |
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. |
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 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 protein. 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.
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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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TAE是使用最广泛的缓冲系统。其特点是超螺旋在其中电泳时更符合实际相对分子质量(TBE中电泳时测出的相对分子质量会大于实际分子质量),且双链线状DNA在其中的迁移率较其他两种缓冲液快约10%,电泳大于13kb的片段时用TAE缓冲液将取得更好的分离效果,此外,回收DNA片段时也易用TAE缓冲系统进行电泳。TAE的缺点是缓冲容量小,长时间电泳(如过夜)不可选用,除非有循环装置使两极的缓冲液得到交换。 50×TAE Buffer 配制方法: 1. 称量Tris 242g,Na2EDTA.2H2O 37.2g 于1L烧杯中; 2.向烧杯中加入约800ml去离子水,充分搅拌均匀; 3加入57.1ml的冰乙酸,充分溶解; 4.加去离子水定容至1L后,室温保存。向左转|向右转
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
此外,我们常常用“电泳法”判断液体的性质,是胶体还是溶液。在高中化学中我们就用过这种方法判断给定的液体是否为胶体。MOPS电泳缓冲液、胚胎干细胞培养生长因子、动物肝细胞分离培养试剂盒、结晶紫细胞群落染色试剂盒、MFN-2蛋白表达检测试剂盒、血红细胞溶解液、磷酸缓冲盐溶液、Hanks平衡盐粉剂、胰蛋白酶溶液、
低离子强度时,迁移率快。但离子强度过低,缓冲液的缓冲容量小,不易维持pH恒定。高离子强度时迁移率慢,因此为了获得更快的反应速度,在缓冲容量允许的范围内,离子强度应该尽可能小。

