Recombinant
RabMAb

Recombinant Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

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Overview

  • Product name

    Anti-Transferrin Receptor antibody [EPR20584]
    See all Transferrin Receptor primary antibodies

  • Description

    Rabbit monoclonal [EPR20584] to Transferrin Receptor

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment within Human Transferrin Receptor aa 100-450. The exact sequence is proprietary.
    Database link: P02786

  • Positive control

    • WB: Human fetal liver and placenta lysates; Mouse spleen, testis and placenta lysates; K562, TF-1, Jurkat and RAW 264.7 whole cell lysates. IHC-P: Human placenta, cerebrum, esophagus and esophageal cancer tissues; Mouse kidney tissue. ICC/IF: K562 and RAW 264.7 cells. IP: K562 whole cell lysate.

  • 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 ab214039 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
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.
WB 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa).

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 [EPR20584] (ab214039)
      Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Lanes 1-2 : Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/2000 dilution
      Lane 3 : Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution

      Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg
      Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg
      Lane 3 : TF-1 (human bone marrow erythroleukemia cell line) whole cell lysate at 20 µg

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

      Predicted band size: 85 kDa
      Observed band size: 90 kDa
      why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 2 seconds; Lane 2: 1 minute; Lane 3: 1 second.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunofluorescent analysis of 100% methanol-fixed RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling Transferrin Receptor with ab214039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing positive staining on RAW 264.7 cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunofluorescent analysis of 100% methanol-fixed K562 (human chronic myelogenous leukemia cell line from bone marrow) cells labeling Transferrin Receptor with ab214039  at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing positive staining on K562 cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Transferrin Receptor with ab214039 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on human placenta (PMID: 27483296). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

    • Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      All lanes : Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Human placenta lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

      Predicted band size: 85 kDa
      Observed band size: 90 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

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

       

    • Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      All lanes : Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution

      Lane 1 : Mouse spleen lysate at 20 µg
      Lane 2 : Mouse testis lysate at 20 µg
      Lane 3 : Mouse placenta lysate at 20 µg
      Lane 4 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

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

      Predicted band size: 85 kDa
      Observed band size: 90 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1/4: 3 seconds; Lane 2: 30 seconds; Lane 3: 1 second.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Transferrin Receptor with ab214039 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on capillaries of human cerebrum (PMID: 6095085). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunohistochemical analysis of paraffin-embedded human esophagus tissue labeling Transferrin Receptor with ab214039 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Weakly cytoplasmic staining on human paracarcinoma esophagus. The staining intensity of human paracarcinoma esophagus was weaker than the human esophageal cancer tissue. Both paracarcinoma and human esophageal cancer tissues have been taken from the same patient (PMID: 24435655). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunohistochemical analysis of paraffin-embedded human esophageal cancer tissue labeling Transferrin Receptor with ab214039 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human esophageal cancer. The staining intensity of human esophageal cancer tissue was stronger than the human paracarcinoma esophagus tissue. Both paracarcinoma and human esophageal cancer tissues have been taken from the same patient (PMID: 24435655). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Transferrin Receptor with ab214039 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on renal tubules of mouse kidney (PMID: 12538733). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

    • Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
      Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)

      Transferrin Receptor was immunoprecipitated from 0.35 mg of K562 (human chronic myelogenous leukemia cell line from bone marrow) lysate with ab214039 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214039 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

      Lane 1: K562 whole cell lysate 10 µg (Input). 

      Lane 2: ab214039 IP in K562 whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214039 in K562 whole cell lysate.

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

      Exposure time: 1 second.

    References

    This product has been referenced in:

    • Huang LL  et al. Augmenter of liver regeneration protects the kidney from ischaemia-reperfusion injury in ferroptosis. J Cell Mol Med 23:4153-4164 (2019). Read more (PubMed: 30993878) »
    See 1 Publication for this product

    Publishing research using ab214039? Please let us know so that we can cite the reference in this datasheet.

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