製品の概要

  • 製品名

    Anti-TSG101 antibody [4A10]
    TSG101 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [4A10] to TSG101
  • 由来種

    Mouse
  • 特異性

    This antibody recognizes the TSG-101 protein, the product of a recently identified tumor susceptibility gene the inactivation of which in mouse fibroblasts results in cell transformation and the ability of those cells to form tumors in nude mice.
  • アプリケーション

    適用あり: Flow Cyt, ICC/IF, IP, WB, IHC-Pmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Fusion protein corresponding to Human TSG101 aa 167-374. Expressed in E.coli
    Database link: Q99816

  • ポジティブ・コントロール

    • WB: Neuro-2a, C8D30, NIH/3T3, RAW 264.7 , C2C12 , HeLa, HepG2, A431, K562 and THP-1 whole cell lysate, mouse testis. ICC/IF: Amyloid peptide-treated mouse astrocytes IHC-P: Colon carcinoma tissue, human ovarian cancer tissue, human breast carcinoma tissue.
  • 特記事項

    This product was changed from ascites to tissue culture supernatant on 12th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab83 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use at an assay dependent concentration. PubMed: 23045692

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/500 - 1/1000.
IP Use at an assay dependent concentration.
WB 1/500 - 1/3000. Detects a band of approximately 47 kDa (predicted molecular weight: 43 kDa).
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 機能

    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.
  • 組織特異性

    Heart, brain, placenta, lung, liver, skeletal, kidney and pancreas.
  • 配列類似性

    Belongs to the ubiquitin-conjugating enzyme family. UEV subfamily.
    Contains 1 SB (steadiness box) domain.
    Contains 1 UEV (ubiquitin E2 variant) domain.
  • ドメイン

    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.
  • 翻訳後修飾

    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.
  • 細胞内局在

    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
  • 参照データベース

  • 別名

    • 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 101 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

画像

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
    Lane 2 : C8D30 whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
    Lane 4 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : C2C12 (mouse myoblast cell line) whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-mouse IgG antibody at 1/500 dilution

    Predicted band size: 43 kDa



    10% SDS PAGE

    Running conditions: 80V for 15min then 140V for 40min

    Blocking: 5% non-fat milk in TBST at room temperature for 60min.

    Washing conditions: 5 ml TBST, 4 x 5min

    Transfer conditions: Semi-dry, 18 V, 60min (NC membrane)

    Exposure system: Trident plus Western HRP Substrate

  • Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling TSG101 with ab83 at 1/100 dilution. Cytoplasmic staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min. (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H2O2, RT, 30min. Blocking condition: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30min. Primary antibody incubation: 4°C overnight. Secondary antibody incubation: HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 minutes. DAB detection. 

  • Immunofluorescence analysis of amyloid peptide-treated mouse astrocytes, staining TSG101 with ab83.

    Astrocytes were fixed with paraformaldehyde and permeabilized with 0.2% Triton X-100 for 5 min at room temperature. Cells were incubated with primary antibody and a fluorescent-conjugated anti-mouse IgG was used as the secondary antibody.
  • Panc-1 and Serum Exosomes were characterized by the exosome-specific expression of TSG101 by FACS analysis.

    Exosomes were attached to 4-μm aldehyde/sulfate latex beads by mixing ∼30 μg of exosomes in a 100-μl volume of beads for 2 hours at room temperature. This suspension was diluted to 1 ml with PBS, and the reaction was stopped with 100 mm glycine and 2% BSA in PBS. Exosome-bound beads were washed in PBS/1% BSA, blocked with 10% BSA, and stained for FACS with anti-TSG101 (1:400, ab83). An Alexa Fluor® 488 conjugated anti-mouse IgG was used as the secondary antibody.

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : Non-transfected (–) 293T whole cell extracts
    Lane 2 : TSG101 shRNA transfected (+) 293T whole cell extracts

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-mouse IgG antibody

    Developed using the ECL technique.

    Predicted band size: 43 kDa



    10% SDS-PAGE

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : Mouse testis tissue extract
    Lane 2 : Mouse pancreas tissue extract

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-mouse IgG antibody

    Developed using the ECL technique.

    Predicted band size: 43 kDa

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling TSG101 with ab83 at 1/100 dilution. Cytoplasmic staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : K562 whole cell extract
    Lane 2 : THP-1 whole cell extract
    Lane 3 : HL-40 whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-mouse IgG antibody

    Developed using the ECL technique.

    Predicted band size: 43 kDa



    10% SDS-PAGE

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : NIH-3T3 whole cell lysate/extract
    Lane 2 : JC (mouse mammary adenocarcinoma) whole cell lysate/extract
    Lane 3 : BCL-1 whole cell lysate/extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-mouse IgG antibody

    Predicted band size: 43 kDa



    10% SDS-PAGE

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

    Lane 1 : A431 whole cell lysate
    Lane 2 : HeLa whole cell lysate
    Lane 3 : HepG2 whole cell lysate
    Lane 4 : K562 whole cell lysate
    Lane 5 : THP-1 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 43 kDa

  • Immunohistochemical analysis of paraffin-embedded colon carcinoma tissue labeling TSG101 with ab83 at 1/100 dilution.

  • All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 whole cell lysate (ab7909)
    Lane 3 : Jurkat whole cell lysate (ab7899)
    Lane 4 : HEK293 whole cell lysate (ab7902)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Donkey polyclonal to Mouse IgG (IRDyeTM 700DX) at 1/10000 dilution

    Predicted band size: 43 kDa
    Observed band size: 49 kDa
    why is the actual band size different from the predicted?

参考文献

This product has been referenced in:

  • Jiao YJ  et al. Characterization and proteomic profiling of pancreatic cancer-derived serum exosomes. J Cell Biochem 120:988-999 (2019). Read more (PubMed: 30160795) »
  • Capello M  et al. Exosomes harbor B cell targets in pancreatic adenocarcinoma and exert decoy function against complement-mediated cytotoxicity. Nat Commun 10:254 (2019). Read more (PubMed: 30651550) »
See all 123 Publications for this product

レビューと Q&A

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1-3 of 3 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Hela
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

投稿 Mar 20 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Human HeLa)
Loading amount
30 µg
Specification
Human HeLa
Gel Running Conditions
Reduced Denaturing (12.5% gel)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

投稿 Nov 25 2009

Application
Western blot
Sample
Rat Tissue lysate - other (brain, HEK cell lysate)
Loading amount
50 µg
Specification
brain, HEK cell lysate
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C

Dr. Sriharsha Kantamneni

Verified customer

投稿 Sep 26 2007

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