Overview

  • Product name
    Anti-TGF alpha antibody [P/T1]
    See all TGF alpha primary antibodies
  • Description
    Mouse monoclonal [P/T1] to TGF alpha
  • Host species
    Mouse
  • Specificity
    This antibody was selected and evaluated immunoserologically for recombinant human TGF-alpha and synthetic (uncoupled) peptides using simple binding and competitive ELISA. Immunohistochemical characterization of the anti-TGF-alpha monoclonal antibody was made on different human carcinomas. Recommended for immunohistochemical detection of TGF-alpha production in human tumor cells, immunoserological monitoring of progression of malignant tumours and for screening of therapeutic efficiency of different human tumors.
  • Tested applications
    Suitable for: ELISA, IHC-P, IHC-Fr, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide fragment (32-40 amino acids sequence of 50 amino acids) of transforming growth factor alpha (TGF-alpha).

Properties

Applications

Our Abpromise guarantee covers the use of ab9578 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
ELISA Use at an assay dependent concentration.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

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

 

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

  • Immunohistochemical characterization of the anti-TGF-a monoclonal antibody ab9578 on human melanoma.
  • Overlay histogram showing A431 cells stained with ab9578 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9578, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Badawy AA  et al. Expression of Epidermal Growth Factor Receptor and Transforming Growth Factor Alpha in Cancer Bladder: Schistosomal and Non-Schistosomal. Curr Urol 9:192-201 (2017). Read more (PubMed: 28413380) »
  • Schrevel M  et al. Autocrine expression of the epidermal growth factor receptor ligand heparin-binding EGF-like growth factor in cervical cancer. Int J Oncol 50:1947-1954 (2017). IHC-P ; Human . Read more (PubMed: 28498437) »
See all 6 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (melanoma)
Specification
melanoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 1.5%

Abcam user community

Verified customer

Submitted Apr 03 2008

Answer

Thank you for contacting your customer and forwarding the responses to us. As I understand, the customer is using A431 whole cell lysate and loaded 10 ug. This is may be not enough for detecting the target protein on Western blot and I would suggest loading at least 25-30 ug protein per lane. It is also important to know that the customer has actually used GST-tagged protein. It may well be that the GST tag somehow blocks the recognition of the antibody. Please let me know how your customer is getting on with 30 ug protein. If there is still a problem, then please do not hesitate to contact us again.

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Answer

Thank you for your enquiry. We are very sorry to hear that your customer is having problem with this antibody. We would like to know some more details of your customer’s Western blot assay. 1. How much protein was loaded onto the gel? 2. Does the customer know the expression levels of TGF-alpha in the samples? Do the A431 cells high or low expression of TGF-alpha? 3. Did the customer check the secondary antibody with another primary to make sure that the detection system works properly? 4. What lysis buffer did the customer specifically use? We would strongly suggest using RIPA buffer. Preparation of RIPA cell lysates for Western Blots: 1. Wash cell pellets once with ice-cold PBS. 2. Add 1 ml of RIPA buffer to 10x8 cells, incubate on ice for 20 min, vortex 2 to 3 times. 3. Centrifuge for 5 min at 4oC at maximum speed in a microfuge tube. 4. Transfer supernatant into clean tube. Measure protein concentration with a protein assay. 5. Adjust concentration to 5 mg/ml with RIPA lysis buffer. 6. Add equal volume of 2 x SDS sample buffer into cell lysate, boil for 5 min. 7. Store at -20oC for daily use or -80oC for long term. Avoid repeated freeze thaw cycles. RIPA Base Ingredients Tris-HCl: 50 mM, pH 7.4 NP-40: 1% Na-deoxycholate: 0.25% NaCl: 150 mM EDTA: 1 mM PMSF: 1 mM Aprotinin, leupeptin, pepstatin: 1 microgram/ml each RIPA Protease Inhibitors Phenylmethylsulfonyl fluoride (PMSF) (200 mM stock solution in isopropanol; store at room temperature) EDTA (calcium chelator; 100 mM stock solution in H2O, pH 7.4) Leupeptin (store frozen in aliquots, 1 mg/ml in H2O) Aprotinin (store frozen in aliquots, 1 mg/ml in H2O) Pepstatin (store frozen in aliquots, 1 mg/ml in methanol) Finally one note, the immunogen used to raise this antibody was a synthetic peptide fragment (32-40 amino acids sequence of 50 amino acids) of transforming growth factor alpha (TGF-alpha). Therefore, we would suggest running a positive control (purified TGF-alpha) along with the samples. We look forward to hearing from you soon.

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Answer

Thank you for your enquiry and for taking your time to fill in the on-line Questionnaire. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. 1. Try to coat the antigen in bicarbonate buffer rather than in PBS: Bicarbonate/carbonate coating buffer (100 mM): 3.03 g Na2CO3, 6.0 g NaHCO3 (1000 ml distilled water) pH 9.6, 2. The coating time should be at least for 2 h at room temperature. 3. We would recommend testing a range 1-10 ug/ml of the antigen in 100 ul of coating buffer. 4. Blocking should be with 5% BSA for at least 2 h at room temperature or, if more convenient, overnight at 4°C. 5. Incubate the wells with the primary antibody for at least 2 hrs. 6. Have you tested the secondary antibody to make sure that detection system works properly? We hope this will help you. Should you still have problem with this antibody or need further assistance then please do not hesitate to contact us again.

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Answer

Thank you for your enquiry and for your patience. Ab8610 recognizes the C-terminal part of TGF-a, as does ab2385. I am still waiting to hear back from the originator of ab9578 abd will contact you once I have.

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Answer

We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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