Anti-Transferrin Receptor antibody [B349 (DF1513)] (ab8598)

Reviews (2)Q&A (11)References (3)

Overview

  • Product name
    Anti-Transferrin Receptor antibody [B349 (DF1513)]
    See all Transferrin Receptor primary antibodies
  • Description
    Mouse monoclonal [B349 (DF1513)] to Transferrin Receptor
  • Host species
    Mouse
  • Specificity
    This antibody reacts with the transferrin receptor, a 180-190 kD transmembrane glycoprotein which exists as a 95 kD homodimer with interchain disulfide bond. The specificity of these antibodies, as demonstrated by immunoprecipitation, are equivalent to OKT9, B3/25 and BerT9. This antibody reacts with many proliferating cells in both normal and neoplastic tissues. It also reacts with renal tubular epithelium, islets of Langerhans, scattered cells in the anterior pituitary, hepatocytes and most tissue macrophages.
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Cell line KG1.

  • Positive control
    • Tonsil.
  • General notes


    This antibody is an indicator of proliferation activity. It also has prognostic significance when typing tumors, such as leukemias and lymphomas.

Properties

Applications

Our Abpromise guarantee covers the use of ab8598 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 1/100.
Flow Cyt Use at an assay dependent concentration.

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

 
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Cellular uptake of iron occurs via receptor-mediated endocytosis of ligand-occupied transferrin receptor into specialized endosomes. Endosomal acidification leads to iron release. The apotransferrin-receptor complex is then recycled to the cell surface with a return to neutral pH and the concomitant loss of affinity of apotransferrin for its receptor. Transferrin receptor is necessary for development of erythrocytes and the nervous system (By similarity). A second ligand, the heditary hemochromatosis protein HFE, competes for binding with transferrin for an overlapping C-terminal binding site. Positively regulates T and B cell proliferation through iron uptake (PubMed:26642240).
    (Microbial infection) Acts as a receptor for new-world arenaviruses: Guanarito, Junin and Machupo virus.
  • Involvement in disease
    Immunodeficiency 46
  • Sequence similarities
    Belongs to the peptidase M28 family. M28B subfamily.
    Contains 1 PA (protease associated) domain.
  • Post-translational
    modifications
    N- and O-glycosylated, phosphorylated and palmitoylated. The serum form is only glycosylated.
    Proteolytically cleaved on Arg-100 to produce the soluble serum form (sTfR).
    Palmitoylated on both Cys-62 and Cys-67. Cys-62 seems to be the major site of palmitoylation.
  • Cellular localization
    Secreted and Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
    • Alternative names
      • CD 71 antibody
      • CD71 antibody
      • CD71 antigen antibody
      • IMD46 antibody
      • OTTHUMP00000208523 antibody
      • OTTHUMP00000208524 antibody
      • OTTHUMP00000208525 antibody
      • p90 antibody
      • sTfR antibody
      • T9 antibody
      • TFR 1 antibody
      • TfR antibody
      • TfR1 antibody
      • TFR1_HUMAN antibody
      • TFRC antibody
      • TR antibody
      • Transferrin receptor (p90 CD71) antibody
      • Transferrin receptor protein 1, serum form antibody
      • Trfr antibody
      see all

    Images

    • Western blot - Anti-Transferrin Receptor antibody [B349 (DF1513)] (ab8598)
      Western blot - Anti-Transferrin Receptor antibody [B349 (DF1513)] (ab8598)This image is courtesy of an anonymous Abreview
      All lanes : Anti-Transferrin Receptor antibody [B349 (DF1513)] (ab8598) at 1/100 dilution

      Lane 1 : Human U2OS cell line, whole cell lysate
      Lane 2 : Human 293T cell line, whole cell lysate
      Lane 3 : African Green Monkey Cos7 cell line, whole cell lysate
      Lane 4 : Hamster CHO cell line, whole cell lysate
      Lane 5 : Rat PC12 cell line, whole cell lysate
      Lane 6 : Mouse 3T3 cell line, whole cell lysate

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : HRP conjugated goat anti-mouse antibody

      Developed using the ECL technique.

      Performed under reducing conditions.

      Exposure time: 7 minutes

      See Abreview

    References

    This product has been referenced in:
    • Perrot G  et al. LRP-1--CD44, a new cell surface complex regulating tumor cell adhesion. Mol Cell Biol 32:3293-307 (2012). Read more (PubMed: 22711991) »
    • Ortiz-Zapater E  et al. Trafficking of the human transferrin receptor in plant cells: effects of tyrphostin A23 and brefeldin A. Plant J 48:757-70 (2006). Read more (PubMed: 17059402) »
    See all 3 Publications for this product

    Publishing research using ab8598? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 13 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Transferrin Receptor antibody [B349 (DF1513)]

     Excellent
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (lung epithelial cells)
    Specification
    lung epithelial cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.2% saponin
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 31 2012

    Question

    3T3 lysates were negative in western blotting but so were 293 cells. We need an antibody positive for human transferrin receptor and negative for mouse.

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

    To check the status of the order please contact our Customer Service team and reference this number.

    Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    Would these products
    (ab1086, ab84036, ab9179, ab8598) detect Trf1 and Trf2?

    Read More
    Answer

    Thank you for your enquiry.

    We have not tested these products (ab1086, ab84036, ab9179) for cross reactivity with Trf2. However, based on immunogen sequence (ClustalW analysis, Score 23.0), we do not expect to see any cross reactivity whatsoever. However, there might be a possibility that ab8598 could react but we do not supporting data.

    I hope this information will help settle any concerns that you may have.

    Read More

    Question

    I recently ordered the Anti-Transferrin Receptor antibody [B349 (DF1513)] (ab8598) from you. I have used it at 1:100, in 1% Milk TBST at 4 oC overnight followed by a secondary anti mouse HRP antibody 1:3000. I got no signal from the antidody, a few weak bands but nothing that I think was specific. Are there any specific conditions that this antibody should be usedunderso I canget it working a little better? Any information will be very much appreciated.

    Read More
    Answer

    Thank you for your inquiry.


    We have confirmedTFR expression in Hep G2, HEL 92.1.7 and TF-1 whole cell lysates using thefollowing conditions:

    Load 50 ug of total protein
    Blocking with 5% milk TBST for one hour at room temperature
    Primary: 1:100 incubate for one hour at room temperature with 5% milk TBST
    Secondary: in-house secondarygoat anti-mouse IgG-HRP: at 1:3000. Incubate for 45 minutes
    10% gelPVDF membranes

    If the above does not help, I can help if you provide more detail on the protocol:

    1. What samples are you testing?
    2. How much protein were you loading?
    3. What did you do for the blocking step?
    4. Have you confirmed the secondary has been performing?
    5. Did you run a loading control?
    6. Did you load a positive control?

    I hope this information helps. Please contact us with any other questions.

    Read More

    Question

    I spoke to you today about the Anti-transferrin receptor antibody (ab1086) that did not work for my cell line (Jurkat). I would like to thank you for offering to send us a replacement.  I would be interested in trying the western blot again with ab8598 (the antibody that you recommended, which seems to have worked quite well with a number of cell lines).   Please let me know if you need any further information from me.

    Read More
    Answer

    Thanks for following up with me. I would be happy to send you a vial of ab8598. I will just need to locate your initial order for ab1086 to process your request. Do you have the purchase order number or Abcam order reference number associated with the purchase of ab1086? Thanks!

    Read More

    Western blot abreview for Anti-Transferrin Receptor antibody [B349 (DF1513)]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS and 293T cell line)
    Loading amount
    30 µg
    Specification
    U2OS and 293T cell line
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 02 2007

    Question

    BATCH NUMBER 128772 ORDER NUMBER ? DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE cell lysate of Caco-2 cells PRIMARY ANTIBODY your anti-TfR antibody, catalog number ab8598, lot 128772, dilution 1:100 = 4 ug/mL. 2 hours room temp, washing etc with TBS/Tween; blocking beforehand with 5% milk in TBS/Tween DETECTION METHOD ECL from Perkin Elmer POSITIVE AND NEGATIVE CONTROLS USED We thought the Caco-2 lysate was going to be our positive control for our subsequent experiments. ANTIBODY STORAGE CONDITIONS 4 degrees SAMPLE PREPARATION cells in a 6 well plate lysed in 100 uL Laemmli sample buffer with 10 mM DTT and protease inhibitor cocktail set V at the recommended concentration. AFter addition of sample buffer, cells were scraped from the bottom with the back of a pipet tip, pipetted up and down in an insulin syringe to make it less viscous, boiled for 10 min at 99 degrees and stored at -20 overnight. AMOUNT OF PROTEIN LOADED 20 uL of the above solution ELECTROPHORESIS/GEL CONDITIONS 10% SDS gel TRANSFER AND BLOCKING CONDITIONS transfer buffer is tris/glycine with methanol, 450 mAmp-hour (45 mAmp for 10 hours in cold room). SECONDARY ANTIBODY Goat anti mouse-HRP, [another company], 1:1000 dilution in blocking buffer HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No ADDITIONAL NOTES Does this antibody only recognize TfR in its native form perhaps? Since in the application notes it sais it can only be used on frozen sections, I'm starting to think that... Please let me know any tips you might have. Thanks.

    Read More
    Answer

    Further to contact with the source of this antibody I have determined that this antibody does not perform well by western blotting. Given that this antibody was detailed as working by western blotting on our datasheets I would like to firstly apolagise and secondly offer you a credit note to the value of one vial of this antibody. Please e-mail me details of your original purchase including the order number and date of purchase and I will arrange for our accounts team to raise you the credit.

    Read More

    Question

    BATCH NUMBER 128772 ORDER NUMBER ? DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE cell lysate of Caco-2 cells PRIMARY ANTIBODY your anti-TfR antibody, catalog number ab8598, lot 128772, dilution 1:100 = 4 ug/mL. 2 hours room temp, washing etc with TBS/Tween; blocking beforehand with 5% milk in TBS/Tween DETECTION METHOD ECL from Perkin Elmer POSITIVE AND NEGATIVE CONTROLS USED We thought the Caco-2 lysate was going to be our positive control for our subsequent experiments. ANTIBODY STORAGE CONDITIONS 4 degrees SAMPLE PREPARATION cells in a 6 well plate lysed in 100 uL Laemmli sample buffer with 10 mM DTT and protease inhibitor cocktail set V at the recommended concentration. AFter addition of sample buffer, cells were scraped from the bottom with the back of a pipet tip, pipetted up and down in an insulin syringe to make it less viscous, boiled for 10 min at 99 degrees and stored at -20 overnight. AMOUNT OF PROTEIN LOADED 20 uL of the above solution ELECTROPHORESIS/GEL CONDITIONS 10% SDS gel TRANSFER AND BLOCKING CONDITIONS transfer buffer is tris/glycine with methanol, 450 mAmp-hour (45 mAmp for 10 hours in cold room). SECONDARY ANTIBODY Goat anti mouse-HRP, [another company], 1:1000 dilution in blocking buffer HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No ADDITIONAL NOTES Does this antibody only recognize TfR in its native form perhaps? Since in the application notes it sais it can only be used on frozen sections, I'm starting to think that... Please let me know any tips you might have. Thanks.

    Read More
    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionnaire and I have a few comments. Whilst we recommend that our customers use a tonsil preparation as a positive control I have performed a brief search of the literature and it appears that the use of Caco-2 cells is quite standard for the studying of Transferrin Receptor expression. I would like to recommend that you quantitate the mass of protein that you are loading onto the gel; presently there is no way of determining the mass of protein loaded per well. This will provide a better idea of whether the antibody is capable of detecting its target by immunoblotting. It would also help if you ponceau stain your membrane following transfer. This will enable you to determine the integrity of the protein and the success of transfer. Alternatively a parallel loading control western blot would serve to confirm the integrity and transfer of your samples. I will submit your question regarding the antibody only targeting the native form of this protein to the lab and get back to you shortly. I appreciate your patience.

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    Question

    Regarding the immunogen, has the epitope been mapped? Is it within the extracellular domain?

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    Answer

    The epitope has not been mapped. The cellular loction is cell membrane.

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    Question

    I have a customer that would like to see a reference or image of this antibody used in IHC. Thanks

    Read More
    Answer

    Thank you for your enquiry. Here are some references: Testa, U., wt al 1993. Crit. Rev. Oncog. 4.241-276, Barclay, A.N., et al. 1997. Leucocyte Antigen FactsBook. Academic Press. London Oudemans, et al. 1986 Cancer 58. 1252. Hentze, M.W. and Kuhn,L.C. 1996. Proc.Natl Acad. Sci. USA 93. 8175-8182 Knapp, W., et al eds. 1989 Leucocyte Typing Workshop IV. Oxford University Press.

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    1-10 of 13 Abreviews or Q&A

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