Overview

  • Product name

    Anti-TGF alpha antibody [EPR15346]
    See all TGF alpha primary antibodies
  • Description

    Rabbit monoclonal [EPR15346] to TGF alpha
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human TGF alpha aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P01135

  • Positive control

    • WB: Human fetal skin, fetal liver and breast cancer lysates; HepG2 and Ramos whole cell lysates. IHC-P: Human liver and hepatocellular carcinoma tissues. ICC/IF: HepG2 cells. IP: HepG2 and Ramos whole cell lysates.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208156 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 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/40.

Target

  • Function

    TGF alpha is a mitogenic polypeptide that is able to bind to the EGF receptor and to act synergistically with TGF beta to promote anchorage-independent cell proliferation in soft agar.
  • Tissue specificity

    Isoform 1, isoform 3 and isoform 4 are expressed in keratinocytes and tumor-derived cell lines.
  • Sequence similarities

    Contains 1 EGF-like domain.
  • Cellular localization

    Cell membrane and Secreted > extracellular space.
  • Information by UniProt
  • Database links

  • Alternative names

    • EGF like TGF antibody
    • EGF-like TGF antibody
    • ETGF antibody
    • TFGA antibody
    • TGF A antibody
    • TGF type 1 antibody
    • TGF-alpha antibody
    • Tgfa antibody
    • TGFA_HUMAN antibody
    • Transforming growth factor alpha antibody
    • Transforming growth factor alpha precursor antibody
    • Wa1 antibody
    • Waved 1 antibody
    see all

Images

  • Lane 1 : Anti-TGF alpha antibody [EPR15346] (ab208156) at 1/1000 dilution
    Lane 2 : Anti-TGF alpha antibody [EPR15346] (ab208156) at 1/5000 dilution

    All lanes : MCF-7 whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 17 kDa
    Observed band size: 15 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    The different result is due to the lysates preparation method. The lysate in the left image is prepared by RIPA lysis method. The lysate in the right image is prepared by 1% SDS Hot lysis method. This antibody works better in 1% SDS Hot Lysates in WB.

    For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling TGF alpha with ab208156 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human liver is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling TGF alpha with ab208156 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody  -Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG  (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab208156 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • TGF alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab208156 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab208156 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HepG2 whole cell lysate 10µg (Input).

    Lane 2: ab208156 IP in HepG2 whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab208156 in HepG2 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

  • All lanes : Anti-TGF alpha antibody [EPR15346] (ab208156) at 1/1000 dilution

    Lane 1 : Human fetal skin lysate
    Lane 2 : Human fetal liver lysate
    Lane 3 : Human breast cancer lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 17 kDa
    Observed band size: 15 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 3: 3 minutes; Lane 2: 1 minute.

  • Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling TGF alpha with ab208156 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on the cancer cells of Human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells, treated with Brefeldin A (10 µM, 18 hours), labeling TGF alpha with ab208156 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cells, treated with Brefeldin A (10 µM, 18 hours). The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG  (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab208156 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • TGF alpha was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell lysate with ab208156 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab208156 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Ramos whole cell lysate, 10µg (Input).

    Lane 2: ab208156 IP in Ramos whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab208156 in Ramos whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • All lanes : Anti-TGF alpha antibody [EPR15346] (ab208156) at 1/1000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 17 kDa
    Observed band size: 15 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 15 seconds; Lane 2: 3  minutes.

References

This product has been referenced in:

  • Zhang G  et al. CD317 Activates EGFR by Regulating Its Association with Lipid Rafts. Cancer Res 79:2220-2231 (2019). Read more (PubMed: 30890618) »
  • Sun L  et al. Decreased expression of acetyl-CoA synthase 2 promotes metastasis and predicts poor prognosis in hepatocellular carcinoma. Cancer Sci 108:1338-1346 (2017). WB, IHC . Read more (PubMed: 28387999) »
See all 2 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab208156.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up