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Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)

Reviews (3)Q&A (8)References (217)
Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
  • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)

Key features and details

  • Mouse monoclonal [4A10] to TSG101 - BSA and Azide free
  • Suitable for: Flow Cyt, WB, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-TSG101 antibody [4A10] - BSA and Azide free
    See all TSG101 primary antibodies

  • Description

    Mouse monoclonal [4A10] to TSG101 - BSA and Azide free

  • Host species

    Mouse

  • Specificity

    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.

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment within Human TSG101 aa 150 to the C-terminus. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. Expressed in E.coli
    Database link: Q99816

  • Positive control

    • 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: human ovarian cancer tissue, human breast carcinoma tissue. Flow cyt: THP-1 cells

  • General notes

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
Flow Cyt
Use at an assay dependent concentration.

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

 
WB (3)
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.
Notes
Flow Cyt
Use at an assay dependent concentration.

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

 
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.

Target

  • Function

    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.

  • Tissue specificity

    Heart, brain, placenta, lung, liver, skeletal, kidney and pancreas.

  • Sequence similarities

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

  • Domain

    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.

  • Post-translational
    modifications

    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.

  • Cellular localization

    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

  • Database links

    • Alternative names

      • 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

    Images

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)

      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. 

    • Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)Image from Kahlert C et al., J Biol Chem. 2014;289(7):3869-75. Fig 1(C).; doi: 10.1074/jbc.C113.532267.

      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.

    • Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Flow Cytometry - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Flow Cytometry analysis of THP-1 cells labeling TSG101 with ab83 at 1/25 dilution (red). Unlabelled sample was used as a control (black). A Dylight 488-conjugated secondary antibody was used for FACS analysis.

       

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (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

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)

      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.

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83) at 1/500 dilution

      Lane 1 : K562 whole cell extract
      Lane 2 : THP-1 whole cell extract
      Lane 3 : HL-60 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

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (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

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (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

    • Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      Western blot - Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83)
      All lanes : Anti-TSG101 antibody [4A10] - BSA and Azide free (ab83) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : A-431 whole cell lysate (ab7909)
      Lane 3 : Jurkat whole cell lysate (ab7899)
      Lane 4 : HEK-293 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?

    References (217)

    Publishing research using ab83? Please let us know so that we can cite the reference in this datasheet.

    ab83 has been referenced in 217 publications.

    • Zhou L  et al. Exosomal LncRNA LINC00659 transferred from cancer-associated fibroblasts promotes colorectal cancer cell progression via miR-342-3p/ANXA2 axis. J Transl Med 19:8 (2021). PubMed: 33407563
    • Matejovic A  et al. Comparison of separation methods for tissue-derived extracellular vesicles in the liver, heart, and skeletal muscle. FEBS Open Bio 11:482-493 (2021). PubMed: 33410274
    • Ke Y  et al. Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90. Stem Cell Res Ther 12:21 (2021). PubMed: 33413616
    • Liu J  et al. Exosomes from tamoxifen-resistant breast cancer cells transmit drug resistance partly by delivering miR-9-5p. Cancer Cell Int 21:55 (2021). PubMed: 33451320
    • Li Z  et al. Exosome-based Ldlr gene therapy for familial hypercholesterolemia in a mouse model. Theranostics 11:2953-2965 (2021). PubMed: 33456582
    View all Publications for this product

    Customer reviews and Q&As

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    1-3 of 3 Abreviews

    Western blot abreview for Anti-TSG101 antibody [4A10]

     Average
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 20 2018

    Western blot abreview for Anti-TSG101 antibody [4A10]

     Good
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 25 2009

    Western blot abreview for Anti-TSG101 antibody [4A10]

     Good
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Sriharsha Kantamneni

    Verified customer

    Submitted Sep 26 2007

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