TSG101 Antibody (4A10)

Novus Biologicals | Catalog # NB200-112

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Canine, Hamster, Monkey, Zebrafish

Cited:

Human, Mouse, Rat, Hamster, Primate, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry Free-Floating, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Electron Microscopy, Knockdown Validated

Cited:

Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Functional Assay, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 4A10
View all TSG101 Primary Antibodies »

Product Specifications

Immunogen

Amino acids 167-374 of TSG101 protein expressed in E. coli.

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Mouse-On-Mouse blocking reagent may be needed for IHC and ICC experiments to reduce high background signal. You can find these reagents under catalog numbers PK-2200-NB and MP-2400-NB. Please contact Technical Support if you have any questions.

Marker

Exosome Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

44 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Whole cell lysates (WCL) or exosome sample from MDA-MB-231 cells was loaded with 10 ug/lane. 10% SDS-PAGE. TSG101 Antibody (NB200-112) was used for primary antibody: 1:1000, 4C, overnight. Image from verified customer review.
Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112]

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112]

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112] - Human ovarian cancer. TSG101 stained by TSG101 antibody [4A10] (NB200-112).Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
Flow Cytometry: TSG101 Antibody (4A10) [NB200-112]

Flow Cytometry: TSG101 Antibody (4A10) [NB200-112]

Flow Cytometry: TSG101 Antibody (4A10) [NB200-112] - TSG101 antibody [4A10] (NB200-112) detects TSG101 protein by flow cytometry analysis.Sample: THP-1 cell.Black: Unlabelled sample was used as a control.Red: TSG101 antibody [4A10] (NB200-112).Acquisition of 20,000 events were collected using a Dylight 488-conjugated secondary antibody for FACS analysis.
Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112] - A. 30 ug NIH-3T3 whole cell lysate/extract. B. 30 ug JC whole cell lysate/extract. C. 30 ug BCL-1 whole cell lysate/extract.
Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with TSG101 antibody [4A10] diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (NBP2-19382) was used to detect the primary antibody.
Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Lane 1: MWM, Lane 2: TSG101 in RAW 264.7 Whole Cell Lysate, Lane 3: Exosome Lysate. Used primary antibody at 1:500 and Donkey Anti-Mouse at 0.5 ug/mL (this could be titrated down to reduce background). Used picogram sensitivity ECL reagents. Image from verified customer review.
Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Various tissue extracts (50 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with TSG101 antibody [4A10] diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (NBP2-19382) was used to detect the primary antibody.
Western Blot: TSG101 Antibody (4A10) [NB200-112]

Western Blot: TSG101 Antibody (4A10) [NB200-112]

TSG101-Antibody-4A10-Western-Blot-NB200-112-img0034.jpg
Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112]

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112]

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112] - Human breast carcinoma. TSG101 stained by TSG101 antibody [4A10] (NB200-112).Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
ELISA: TSG101 Antibody (4A10) [NB200-112]

ELISA: TSG101 Antibody (4A10) [NB200-112]

ELISA: TSG101 Antibody (4A10) [NB200-112] - ELISA detection of TSG101 for capture at a concentration of 5 ug/mL and TSG101 for detection at a concentration of 1.5 ug/mL.
Knockdown Validated: TSG101 Antibody (4A10) [NB200-112]

Knockdown Validated: TSG101 Antibody (4A10) [NB200-112]

Knockdown Validated: TSG101 Antibody (4A10) [NB200-112] - Non-transfected (-) and transfected (+) 293T whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with TSG101 antibody [4A10] diluted at 1:500.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Characterization of pECs-EVs from different women in secretory & proliferative phase of cycle. Representative TEM images of pECs-EVs from secretory (A,B) & proliferative phase (C,D). Scale bars: 50 nm (A,C); 100 nm (B,D). Representative nanoparticle tracking analysis plots of pECs-EVs from secretory (E) & proliferative phase (F). Western blot showing the presence of different canonical EV markers (CD63, CD9, Alix & TSG101) in secretory & proliferative phase pECs-EVs (n = 4) (G). Negative EV markers used were Calnexin (ER), beta -tubulin & the cytosolic form of DCXR (H). The same markers were evaluated in pECs protein extract as controls for EV enrichment of these markers. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32483153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Characterization of exosomes. (A) Representative TEM images of exosomes at 22000x magnification (Left) HCT116-R exosomes & (Right) HCT116-P exosomes. (B) Enlarged planar view of a cup-shaped structure for single exosome in our samples at 87000x magnification. (C) Western blot analysis of exosomes revealed differential expression of TSG101, CD9, & CD63 between HCT116-R exosomes & HCT116-P exosomes. (D) Average size distribution & zeta potential of exosomes measured in Zetasizer. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31712601), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Western Blot: TSG101 Antibody (4A10) [NB200-112] - EPS15 depletion affects late endosomal maturation.(A) Co-depletion of SPOPL an EPS15 has an additive effect on influenza A virus infection. (B) EPS15 depletion stabilizes ESCRT components HRS, STAM & TSG101. (C) EPS15 depletion affects LDL uptake in cells resulting in an accumulation of LDL in enlarged vacuoles (upper panel). Late endosomes, visualized by life-cell microscopy of GFP-RAB7, are enlarged in cells depleted of EPS15 (lower panel).DOI:http://dx.doi.org/10.7554/eLife.13841.016 Image collected & cropped by CiteAb from the following publication (https://elifesciences.org/articles/13841), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112] -

Immunohistochemistry-Paraffin: TSG101 Antibody (4A10) [NB200-112] - TSG101 antibody [4A10] detects TSG101 protein at cytoplasm by immunohistochemical analysis.
Sample: Paraffin-embedded human lung cancer.
TSG101 stained by TSG101 antibody [4A10] (NB200-112) diluted at 1:50.
Antigen Retrieval: Citrate buffer, pH 6.0, 15 min
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with TSG101 antibody [4A10] (NB200-112) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Western Blot: TSG101 Antibody (4A10) [NB200-112] - Various whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with TSG101 antibody [4A10] (NB200-112) diluted at 1:500. The HRP-conjugated anti-mouset IgG antibody was used to detect the primary antibody.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

(A) Schematic representation of the tetraspanin-based recombinant proteins composition. CD9, CD63 and CD81 tetraspanins are fused to RFP reporter protein, carrying on its 5’terminus a cathepsin B-specific active cleavage site (CS) or non-active cleavage site (NACS). (B) Schematic representation showing the expected tetraspanin-based recombinant proteins orientation on EV membrane and cathepsin B-dependent RFP release. (C) Western blot analysis of cathepsin B from 5637, HEK293 and HeLa cells and respective EVs, representative of three independent experiments; detection of beta-actin for cell extracts and TSG101 for EV extracts was used as loading controls. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36579638), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Exogenous CD9/CD81-RFP CS/NACS protein expression in HEK293 cell lysates and EV fraction. A Fluorescence microscopy images of HEK293 cells stably expressing CD9/CD81-RFP CS/NACS recombinant proteins. Representative western blot analysis comparison between CD9 (B) or CD81 (D) from CD9/CD81-RFP CS/NACS HEK293 cells lysates versus EVs. C, Representative western blot analysis comparison of CD81 and CD63 from CD9-RFP CS/NACS HEK293 derived EVs (C) versus CD9 and CD63 from CD81-RFP CS/NACS HEK293 derived EVs (E). The CD9/CD81-RFP fusion protein was detected also with anti-RFP antibody (CD9-RFP in C e CD81-RFP in E). Alix and TSG101 analysis was used for quantitative protein normalization and RFP detection to confirm the presence of recombinant proteins. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36579638), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Exogenous CD9/CD81-RFP CS/NACS protein expression in HEK293 cell lysates and EV fraction. A Fluorescence microscopy images of HEK293 cells stably expressing CD9/CD81-RFP CS/NACS recombinant proteins. Representative western blot analysis comparison between CD9 (B) or CD81 (D) from CD9/CD81-RFP CS/NACS HEK293 cells lysates versus EVs. C, Representative western blot analysis comparison of CD81 and CD63 from CD9-RFP CS/NACS HEK293 derived EVs (C) versus CD9 and CD63 from CD81-RFP CS/NACS HEK293 derived EVs (E). The CD9/CD81-RFP fusion protein was detected also with anti-RFP antibody (CD9-RFP in C e CD81-RFP in E). Alix and TSG101 analysis was used for quantitative protein normalization and RFP detection to confirm the presence of recombinant proteins. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36579638), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

EPS15 depletion affects late endosomal maturation.(A) Co-depletion of SPOPL an EPS15 has an additive effect on influenza A virus infection. (B) EPS15 depletion stabilizes ESCRT components HRS, STAM and TSG101. (C) EPS15 depletion affects LDL uptake in cells resulting in an accumulation of LDL in enlarged vacuoles (upper panel). Late endosomes, visualized by life-cell microscopy of GFP-RAB7, are enlarged in cells depleted of EPS15 (lower panel).DOI:http://dx.doi.org/10.7554/eLife.13841.016 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27008177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Characterization of urine and urinary exosomes isolated by BEST of PE and HC. (A) Representative nanoparticle tracking analysis for untreated urine and isolated urine from PE and HC, depicting particle diameter and the number of particles. (B) and (C) Comparisons of particle concentration and particle size between PE and HC. (D) Western blots of TSG101 and CD63 protein extracted from isolated exosomes. (E) and (F) Quantitative analysis of exosome marker TSG101 and CD63 protein. All data are presented as mean \documentclass[12pt]{minimal}\usepackage{amsmath}\usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy}\usepackage{mathrsfs}\usepackage{upgreek}\setlength{\oddsidemargin}{-69pt}\begin{document}$$\:\pm\:$$\end{document} standard error and analyzed by paired t-test and independent two-tailed t-test. BEST, biologically intact exosome separation technology; PE, patients with preeclampsia; HC, healthy controls. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39406891), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Characterization of exosomes. (A) Representative TEM images of exosomes at 22000x magnification (Left) HCT116-R exosomes and (Right) HCT116-P exosomes. (B) Enlarged planar view of a cup-shaped structure for single exosome in our samples at 87000x magnification. (C) Western blot analysis of exosomes revealed differential expression of TSG101, CD9, and CD63 between HCT116-R exosomes and HCT116-P exosomes. (D) Average size distribution and zeta potential of exosomes measured in Zetasizer. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31712601), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TSG101 Antibody (4A10)

Western Blot: TSG101 Antibody (4A10) [NB200-112] -

Characterization of pECs-EVs from different women in secretory and proliferative phase of cycle. Representative TEM images of pECs-EVs from secretory (A,B) and proliferative phase (C,D). Scale bars: 50 nm (A,C); 100 nm (B,D). Representative nanoparticle tracking analysis plots of pECs-EVs from secretory (E) and proliferative phase (F). Western blot showing the presence of different canonical EV markers (CD63, CD9, Alix and TSG101) in secretory and proliferative phase pECs-EVs (n = 4) (G). Negative EV markers used were Calnexin (ER), beta -tubulin and the cytosolic form of DCXR (H). The same markers were evaluated in pECs protein extract as controls for EV enrichment of these markers. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32483153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TSG101 Antibody (4A10)

Application
Recommended Usage

ELISA

1:100 - 1:2000

Electron Microscopy

Assay dependent

Flow Cytometry

1:25-1:200

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:100-1:1000

Immunohistochemistry Free-Floating

Assay dependent

Immunohistochemistry-Paraffin

1:100-1:1000

Immunoprecipitation

Assay dependent

Western Blot

1:500-1:3000

Reviewed Applications

Read 3 reviews rated 4.3 using NB200-112 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

PBS

Preservative

No Preservative

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TSG101

Tumor susceptibility 101 protein (human TSG101 theoretical molecular weight 44kDa) is the mammalian homologue of the yeast protein Vps23 which plays a role in endosomal and multivesicular body trafficking. TSG101 forms part of the endosomal sorting complexes required for transport complex 1 (ESCRT-I), a cytosolic multiprotein complex consisting additionally of Vps28, Vps37 (A-D) and Mvb12 (A, B) or UBAP1 (1). TSG101 contains multiple domains including the amino terminal ubiquitin e2 variant (UEV) domain, a proline-rich (RRR) domain, a coiled coil (CC) domain, and a carboxy terminal alpha-helical/steadiness box (SB) domain (2). As part of the ESCRT-I complex, TSG101 interacts through its UEV domain with ubiquitinated membrane proteins and members of the ESCRT-0 complex. The UEV domain plays a critical role for various TSG101 functions such as protein sorting into multivesicular bodies and late endosomes, and in the process of viral budding. For example, TSG101 interacts with the intermediate-conductance, Ca2+ -activated K+ channel (KCa3.1), facilitating its targeting to the lysosome for degradation (3). Additionally, TSG101 has been implicated in the turnover of connexins such as connexins 43 and 45 (4). TSG101 plays a role in other cellular functions including transcriptional regulation, cytokinesis and cell growth (2).

Upon its initial discovery, TSG101 was recognized as a tumor suppressor protein due to the identification of deletions within the TSG101 gene in human breast carcinomas. However, re-examination of the initial findings argued against this function and supported that TSG101 promotes tumorigenesis (2). In agreement with this role, TSG101 expression is upregulated in several types of cancer including breast, ovarian, and colorectal carcinoma.

References

1. Schmidt, O., & Teis, D. (2012). The ESCRT machinery. Current Biology. https://doi.org/10.1016/j.cub.2012.01.028

2. Jiang, Y., Ou, Y., & Cheng, X. (2013). Role of TSG101 in cancer. Frontiers in Bioscience. https://doi.org/10.2741/4099

3. Balut, C. M., Gao, Y., Murray, S. A., Thibodeau, P. H., & Devor, D. C. (2010). ESCRT-dependent targeting of plasma membrane localized KCa3.1 to the lysosomes. American Journal of Physiology - Cell Physiology. https://doi.org/10.1152/ajpcell.00120.2010

4. Su, V., & Lau, A. F. (2014). Connexins: Mechanisms regulating protein levels and intercellular communication. FEBS Letters. https://doi.org/10.1016/j.febslet.2014.01.013

Long Name

Tumor Susceptibility 101

Alternate Names

TSG10, VPS23

Entrez Gene IDs

7251 (Human); 22088 (Mouse)

Gene Symbol

TSG101

UniProt

Q99816 (Human)

Additional TSG101 Products

Product Documents for TSG101 Antibody (4A10)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for TSG101 Antibody (4A10)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for TSG101 Antibody (4A10)

Customer Reviews for TSG101 Antibody (4A10) (3)

4.3 out of 5
3 Customer Ratings
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Showing  1 - 3 of 3 reviews Showing All
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  • TSG101 Antibody (4A10)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MDA-MB-231
    Species: Human
    Verified Customer | Posted 10/20/2022
    Western Blot: whole cell lysates (WCL) or exosome sample from MDA-MB-231 cells was loaded with 10 ug/lane. 10% SDS-PAGE. TSG101 Antibody (NB200-112) was used for primary antibody: 1:1000, 4℃, overnight.
    TSG101 Antibody (4A10) NB200-112
  • TSG101 Antibody (4A10)
    Name: Elizabeth Soberg
    Application: Western Blot
    Sample Tested: RAW 264.7 mouse monocyte/macrophage cell line
    Species: Mouse
    Verified Customer | Posted 06/05/2018
    TSG101 in RAW 264.7 Whole Cell Lysate and Exosome Lysate
    Used primary @ 1:500 and Donkey Anti-Mouse @ 0.5 ug/mL (this could be titrated down to reduce background). Used picogram sensitivity ECL reagents.
    TSG101 Antibody (4A10) NB200-112
  • TSG101 Antibody (4A10)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Primary mouse hepatocytes
    Species: Mouse
    Verified Customer | Posted 12/08/2017
    mouse hepatocytes were treated with 100 ng/ml LPS for 4 hours
    mouse hepatocytes were treated with 100 ng/ml LPS for 4 hours
    TSG101 Antibody (4A10) NB200-112

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