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  1. Link

    tsg101-antibody-ab30871.pdf

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Anti-TSG101 antibody (ab30871)

  • Datasheet
  • SDS
Reviews (4)Q&A (4)References (124)

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Western blot - Anti-TSG101 antibody (ab30871)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody (ab30871)
  • Immunocytochemistry/ Immunofluorescence - Anti-TSG101 antibody (ab30871)
  • Western blot - Anti-TSG101 antibody (ab30871)

Key features and details

  • Rabbit polyclonal to TSG101
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-TSG101 antibody [EPR7130(B)] (ab125011)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-TSG101 antibody
    See all TSG101 primary antibodies
  • Description

    Rabbit polyclonal to TSG101
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human TSG101 aa 350 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    Database link: Q99816
    (Peptide available as ab30870)

  • Positive control

    • 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.
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Cycle Inhibitors
    • Other
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Differentiation
    • Cancer
    • Cell cycle
    • Cell differentiation
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • TSG101 peptide (ab30870)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
WB (1)
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 (1)
Use a concentration of 5 µg/ml.
IHC-P (1)
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
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.

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.
  • Target information above from: UniProt accession Q99816 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7251 Human
    • Entrez Gene: 22088 Mouse
    • Entrez Gene: 292925 Rat
    • Omim: 601387 Human
    • SwissProt: Q99816 Human
    • SwissProt: Q61187 Mouse
    • SwissProt: Q6IRE4 Rat
    • Unigene: 523512 Human
    • Unigene: 241334 Mouse
    • Unigene: 7410 Rat
    see all
  • 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 (ab30871)
    Western blot - Anti-TSG101 antibody (ab30871)
    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
    All lanes : 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?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody (ab30871)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody (ab30871)
    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.
  • Immunocytochemistry/ Immunofluorescence - Anti-TSG101 antibody (ab30871)
    Immunocytochemistry/ Immunofluorescence - Anti-TSG101 antibody (ab30871)

    ab30871 stained in HeLa cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab30871 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Western blot - Anti-TSG101 antibody (ab30871)
    Western blot - Anti-TSG101 antibody (ab30871)
    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
    All lanes : 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.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (124)

Publishing research using ab30871? Please let us know so that we can cite the reference in this datasheet.

ab30871 has been referenced in 124 publications.

  • Romano R  et al. Alteration of the late endocytic pathway in Charcot-Marie-Tooth type 2B disease. Cell Mol Life Sci 78:351-372 (2021). PubMed: 32280996
  • Baris IC  et al. Expression and DNA methylation profiles of EZH2-target genes in plasma exosomes and matched primary tumor tissues of the patients with diffuse large B-cell lymphoma. Clin Transl Oncol 23:1152-1166 (2021). PubMed: 33226554
  • Liu X  et al. Hypoxic bone marrow mesenchymal cell-extracellular vesicles containing miR-328-3p promote lung cancer progression via the NF2-mediated Hippo axis. J Cell Mol Med 25:96-109 (2021). PubMed: 33219752
  • Ma S  et al. Gain-of-function p53 protein transferred via small extracellular vesicles promotes conversion of fibroblasts to a cancer-associated phenotype. Cell Rep 34:108726 (2021). PubMed: 33567287
  • Nguyen SL  et al. Integrins mediate placental extracellular vesicle trafficking to lung and liver in vivo. Sci Rep 11:4217 (2021). PubMed: 33602965
View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Immunocytochemistry/ Immunofluorescence abreview for Anti-TSG101 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Kidney)
Permeabilization
Yes - 0.1% Triton X-100
Specification
Kidney
Blocking step
Serum as blocking agent for 50 minute(s) · Concentration: 10% · Temperature: 30°C
Fixative
Formaldehyde
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 26 2020

Immunohistochemistry (Frozen sections) abreview for Anti-TSG101 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (colon)
Permeabilization
No
Specification
colon
Blocking step
Ventanas blocking kit as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 30°C
Fixative
Formaldehyde
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 26 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TSG101 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Venta cell conditioner
Permeabilization
No
Specification
liver
Blocking step
Ventanas blocking kit as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Formaldehyde
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 2020

Western blot abreview for Anti-TSG101 antibody

Excellent
Abreviews
Abreviews
abreview image
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
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 Oct 03 2008

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