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Anti-TSG101 抗体 (ab30871)

Reviews (4)Q&A (4)References (191)
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

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-TSG101 antibody [EPR7130(B)] (ab125011)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-TSG101 antibody
    TSG101 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to TSG101

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, ICC/IF, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    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)

  • ポジティブ･コントロール

    • 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.

  • 特記事項

    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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab30871の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
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.
特記事項
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.

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

    see all

  • 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

  • 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.

参考文献 (191)

ab30871 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab30871 は 191 報の論文で使用されています。

  • Nalbach K  et al. Spatial proteomics reveals secretory pathway disturbances caused by neuropathy-associated TECPR2. Nat Commun 14:870 (2023). PubMed: 36797266
  • Chen X  et al. M1 Microglia-derived Exosomes Promote Activation of Resting Microglia and Amplifies Proangiogenic Effects through Irf1/miR-155-5p/Socs1 Axis in the Retina. Int J Biol Sci 19:1791-1812 (2023). PubMed: 37063422
  • Fratantonio D  et al. The RNA cargo in small extracellular vesicles from chicken eggs is bioactive in C57BL/6 J mice and human peripheral blood mononuclear cells ex vivo. Front Nutr 10:1162679 (2023). PubMed: 37305095
  • Jiang X  et al. Plasma exosomes and contained MiRNAs affect the reproductive phenotype in polycystic ovary syndrome. FASEB J 37:e22960 (2023). PubMed: 37335566
  • Zheng W  et al. Identification of scaffold proteins for improved endogenous engineering of extracellular vesicles. Nat Commun 14:4734 (2023). PubMed: 37550290
View all Publications for this product

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1-8 of 8 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-TSG101 antibody

 Good
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

投稿 Nov 26 2020

Immunohistochemistry (Frozen sections) abreview for Anti-TSG101 antibody

 Good
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

投稿 Nov 26 2020

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

 Good
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

投稿 Nov 25 2020

Western blot abreview for Anti-TSG101 antibody

 Excellent
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

投稿 Oct 03 2008

Question


Responses to your questions as per below:

1) positive controls are of suspected exosomes, which showed the bands at about 26-27 kDa. Negative controls of cells and cell debris showed nothing. Cells collected at 300 x g, cells and cell debris collected at 165000 x g. Exosomes collected after overnight spin @ 120000 x g. Protein extraction performed w/ MPER & quantitated w/ BCA protein assay.

2) Have not tested any other cell types yet.

3) Initially tested tapa1 (cd81) in the N2a cell exosomes but was not working; literature does suggest tsg101 is prevalent in N2a exosomes.

Read More
Answer



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 running a western blot with one of these or another positive control sample.

If one of these sample types is not available, please note we do sell tissue extracts ready to load on a SDS- PAGE gel, such as:

https://www.abcam.com/Liver-Mouse-Whole-Cell-Lysate-normal-tissue-ab29301.html

Read More

Question

Inquiry: Hello, I am using the above stated antibody to detect for exosomes in cells (mouse). 50ug of protein was loaded for every sample and run on a 10% BIS-TRIS gel, w/ transfer using an iBLOT. It was blocked w/ TBS-T-1%BSA, then incubated at 4 degrees overnight with tsg101 (at 1ug/ml, or a 1:600 dilution of the provided 0.6 mg/ml Ab in TBS-T). Washes were performed w/ TBS-T prior to secondary Ab application of goat anti-rabbit at 1:10000 dilution in TBS-T for 1 hour. Washed with TBS, then applied chemiluminescence (Supersignal Pico kit - Pierce), and exposed. I am detecting bands in the desired lanes, although it is at the incorrect size. I got bands around 26-27kDa, rather than at 44 kDa. Is there a problem w/ the lot of antibody, have others come across this, or is it just a fluke and possibly my sample? Thank you for your assistance.

Read More
Answer

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, including method of preparation.

2. Have you prepared any of the cell types shown on datasheet to express detectable levels of this protein.

Read More

Question

Good morning
Do you know if there will be any problem to detect by IF Tsg101 in Vero cells (monkey) using anti Tsg101 rabbit polyclonal antibody (ab30871)

Thank you very much

Read More
Answer

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 in ICC/IF ( with human, mouse and rat).

Therefore, I can offer a discount off a future purchase if you buy ab30871 now, test it in african green monkey and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab30871 in african green monkey. I will then send a discount code. This code must be issued before purchasing ab30871 so please wait for my reply before ordering.

2. Purchase ab30871 either by phone, fax, or online (www.abcam.com).

3. Test it in african green monkey.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab30871 turns out to be unsuitable for african green monkey, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Question

DESCRIPTION OF THE PROBLEM Multiple bands and weak bands SAMPLE Human/ HeLa cell line extract PRIMARY ANTIBODY the purchased one 1:250 and 1:500 in PBS-T (1% Tween-20) over night @ 4°C washing: 1 x 10 min in PBS-T with 5% milk, 2 x 10min in PBS-T DETECTION METHOD ECL and ECl plus POSITIVE AND NEGATIVE CONTROLS USED none ANTIBODY STORAGE CONDITIONS Aliquots at -20°C SAMPLE PREPARATION Laemmli Buffer WITH b-mercaptoethanol 5 min at 95°C AMOUNT OF PROTEIN LOADED 10, 30 and 60 ug ELECTROPHORESIS/GEL CONDITIONS Reducing, 4-12% gradient gel TRANSFER AND BLOCKING CONDITIONS Transfer: semidry, 1 h at 20V Blocking: 5% Milk in PBS-T, 1 h at RT SECONDARY ANTIBODY HRP-Rabbit (works with all other rabbit poly- and monoclonal primary antibodies in our laboratory) Incubation at RT for 1 h same washing step like after primary antibody HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Dilution factor of antibody Amount of cell lysate loaded

Read More
Answer

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 your results. Regarding your samples - how are the lysates prepared? What buffer and protease inhibitors do you include? Can you provide an image of your results? Are you detecting a band around the expected molecular weight or are all bands non-specific and faint? For the primary antibody incubation, it is important to include a blocking reagent in the diluent to help reduce non-specific bands. I recommend diluting the antibody in 5% milk in TBST. Additionally, it is not necessary to include milk in the washes and you can perform washes with TBST alone. TBS is recommended instead of PBS here to help reduce background as well, but is not absolutely necessary to change. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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