概述

  • 产品名称Anti-TSG101抗体
    参阅全部 TSG101 一抗
  • 描述
    兔多克隆抗体to TSG101
  • 经测试应用适用于: WB, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human TSG101.

    (Peptide available as ab30870.)

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) A431 (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) HEK293 (Human embryonic kidney cell line) NIH 3T3 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) This antibody also gave a positive signal in human placenta tissue sections.

性能

应用

Our Abpromise guarantee covers the use of ab30871 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48,49 kDa (predicted molecular weight: 44 kDa).Can be blocked with TSG101 peptide (ab30870).

The doublet seen in Western blot has been described in PMID:11427703 and may be due to internal initiation at Met10.

ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能Component of the ESCRT-I complex, a regulator of vesicular trafficking process. Binds to ubiquitinated cargo proteins and is required for the sorting of endocytic ubiquitinated cargos into multivesicular bodies (MVBs). Mediates the association between the ESCRT-0 and ESCRT-I complex. Required for completion of cytokinesis; the function requires CEP55. May be involved in cell growth and differentiation. Acts as a negative growth regulator. Involved in the budding of many viruses through an interaction with viral proteins that contain a late-budding motif P-[ST]-A-P. This interaction is essential for viral particle budding of numerous retroviruses.
  • 组织特异性Heart, brain, placenta, lung, liver, skeletal, kidney and pancreas.
  • 序列相似性Belongs to the ubiquitin-conjugating enzyme family. UEV subfamily.
    Contains 1 SB (steadiness box) domain.
    Contains 1 UEV (ubiquitin E2 variant) domain.
  • 结构域The UEV domain is required for the interaction of the complex with ubiquitin. It also mediates the interaction with PTAP/PSAP motifs of HIV-1 P6 protein and human spumaretrovirus Gag protein.
    The coiled coil domain may interact with stathmin.
    The UEV domain binds ubiquitin and P-[ST]-A-P peptide motif independently.
  • 翻译后修饰Monoubiquitinated at multiple sites by LRSAM1 and by MGRN1. Ubiquitination inactivates it, possibly by regulating its shuttling between an active membrane-bound protein and an inactive soluble form. Ubiquitination by MGRN1 requires the presence of UBE2D1.
  • 细胞定位Cytoplasm. Membrane. Nucleus. Late endosome membrane. Mainly cytoplasmic. Membrane-associated when active and soluble when inactive. Depending on the stage of the cell cycle, detected in the nucleus. Colocalized with CEP55 in the midbody during cytokinesis.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ESCRT I complex subunit TSG101 antibody
    • ESCRT-I complex subunit TSG101 antibody
    • TS101_HUMAN antibody
    • TSG 10 antibody
    • TSG 101 antibody
    • TSG10 antibody
    • Tsg101 antibody
    • Tumor susceptibility gene 10 antibody
    • Tumor susceptibility gene 101 antibody
    • Tumor susceptibility gene 101 protein antibody
    • Tumor susceptibility protein antibody
    • Tumor susceptibility protein isoform 3 antibody
    • VPS 23 antibody
    • VPS23 antibody
    see all

Anti-TSG101 antibody 图像

  • All lanes : Anti-TSG101 antibody (ab30871) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 44 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)

    See Abreview

  • All lanes : Anti-TSG101 antibody (ab30871) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 44 kDa
    Observed band size : 48,49 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 29 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 90 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab30871 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

    The doublet seen in Western blot has been described in PMID:11427703 and may be due to internal initiation at Met10.

  • ICC/IF image of ab30871 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab30871, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HepG2 and MCF7 cells.

  • IHC image of ab30871 staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab30871, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-TSG101 antibody (ab30871)参考文献

This product has been referenced in:
  • Wiklander OP  et al. Extracellular vesicle in vivo biodistribution is determined by cell source, route of administration and targeting. J Extracell Vesicles 4:26316 (2015). Read more (PubMed: 25899407) »
  • Svensson KJ  et al. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J Biol Chem 288:17713-24 (2013). Read more (PubMed: 23653359) »

See all 7 Publications for this product

Product Wall



I think it would be helpful to have a positive control sample in this instance. The online data for ab30871 shows a western blot where tissue extracts of mouse and rat liver and kidney exhibit detectable levels of TSG101. I would recommend ru...

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Thanks for your enquiry.

Just a few additional questions:

1. Younote you"found bands in thedesired lanes". Are there negative and positive controls in your experiemtn? If so, please briefly describe these controls samples,...

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Thank you very much for your interest in ab30871.

To our knowledge, ab30871 has not been tested in african green monkey and we do not know yet if this antibody will work with Vero cells. I can confirm ab30871 is tested and guaranteed to work...

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Thank you for contacting Abcam regarding ab30871. I am sorry that you have been experiencing difficulties with this antibody in WB. I have reviewed the protocol information you have provided and would like to make some suggestions that may improve ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 20000 cells
Specification HeLa
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Oct 03 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"