Anti-Transferrin Receptor antibody [EPR20584] ab214039 is a rabbit monoclonal antibody that is used in Transferrin Receptor western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20584 is cited in over 90 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | WB | ICC/IF | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Tested | Tested |
Rat | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Cellular uptake of iron occurs via receptor-mediated endocytosis of ligand-occupied transferrin receptor into specialized endosomes (PubMed:26214738). 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 hereditary 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). Acts as a lipid sensor that regulates mitochondrial fusion by regulating activation of the JNK pathway (PubMed:26214738). When dietary levels of stearate (C18:0) are low, promotes activation of the JNK pathway, resulting in HUWE1-mediated ubiquitination and subsequent degradation of the mitofusin MFN2 and inhibition of mitochondrial fusion (PubMed:26214738). When dietary levels of stearate (C18:0) are high, TFRC stearoylation inhibits activation of the JNK pathway and thus degradation of the mitofusin MFN2 (PubMed:26214738). Mediates uptake of NICOL1 into fibroblasts where it may regulate extracellular matrix production (By similarity). (Microbial infection) Acts as a receptor for new-world arenaviruses: Guanarito, Junin and Machupo virus.
CD71, Transferrin receptor protein 1, TR, TfR, TfR1, Trfr, T9, p90, TFRC
Anti-Transferrin Receptor antibody [EPR20584] ab214039 is a rabbit monoclonal antibody that is used in Transferrin Receptor western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20584 is cited in over 90 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Anti-Transferrin Receptor antibody [EPR20584] (ab214039) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P, IP and WB.
Anti-Transferrin Receptor antibody [EPR20584] (ab214039) was first used in a scientific publication in 2019 and has been cited over 52 times in peer reviewed journals. It's performance in Western blot in human and mouse samples is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-Transferrin Receptor antibody [EPR20584] (ab214039) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-Transferrin Receptor antibody [EPR20584] (ab214039) specifically detects Transferrin Receptor (UniProt ID: P02786; Molecular weight: 85kDa) and is sold in 100 µL and 1 mL selling sizes.
Conjugation-ready, carrier free format available for antibody clone EPR20584 - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free ab232376.
Antibody clone EPR20584 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488 (Alexa Fluor® 488 Anti-Transferrin Receptor antibody [EPR20584] ab317529).
The transferrin receptor, crucial for cellular iron uptake, is often overexpressed in cancer cells, making it a valuable target for antibody-mediated therapies. Its role in promoting tumor growth and survival highlights its potential as a therapeutic target and a prognostic marker in oncology.
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 monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
The transferrin receptor commonly referred to as TfR or CD71 is an integral membrane protein that facilitates the uptake of transferrin-bound iron into cells. This receptor has a molecular weight of around 95 kDa and often exists as a homodimer on the cell surface. It is widely expressed in many tissues especially in erythroid precursors and rapidly dividing cells. Alternate names for this receptor include TfR1 and TfR2 though they have distinct roles and distributions. Other transmembrane proteins like OX26 and MEM have been studied in relation to the transferrin receptor due to their involvement in drug delivery.
TfR plays a critical role in iron homeostasis by mediating the internalization of transferrin and release of iron in the endosomes. It operates as part of the transferrin-transferrin receptor complex facilitating iron assimilation necessary for DNA synthesis and cell growth. Iron release involves acidifying endosomes allowing transferrin to bind with specific cellular receptors including alternate forms like beta 2 transferrin. The process subsequently contributes to erythropoiesis and various metabolic processes by regulating essential cellular iron levels.
The transferrin receptor is central to iron metabolism and the receptor-mediated endocytosis pathway. It tightly interacts with transferrin and intracellular pathways process the iron released from transferrin within endosomes. The receptor's role in this pathway involves a dynamic with other proteins such as HFE and hepcidin. These interactions help control systemic iron levels linking closely to the maintenance of erythroid cell health and proliferation.
Disruptions in transferrin receptor function correlate with anemia and neurodegenerative disorders. In anemia related to iron deficiency impaired TfR activity reduces iron uptake culminating in insufficient erythropoiesis. Altered receptor expression or function also connects to neurological diseases like Alzheimer's where iron dysregulation is a concern. Here the transferrin receptor interacts with proteins like Amyloid precursor protein contributing to disease pathology through improper metal homeostasis.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab214039 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
All lanes: Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] (ab214039)
Predicted band size: 84 kDa
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.
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.
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.
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.
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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
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.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 2 seconds; Lane 2: 1 minute; Lane 3: 1 second.
Lanes 1 - 2: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/2000 dilution
Lane 3: Western blot - 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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 seconds; Lane 2: 15 seconds.
All lanes: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution
Lane 1: Human fetal liver lysate at 20 µg
Lane 2: Human placenta lysate at 20 µg
All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/4000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/4: 3 seconds; Lane 2: 30 seconds; Lane 3: 1 second.
All lanes: Western blot - 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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Exposure Time: Lane 1-4: 3 seconds, Lane 5-6: 20 seconds.
Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) used.
Soluble transferrin receptor (sTfR) is cleaved from whole TfR at Arg 100. (PMID: 9575164, PMID: 23390091).
All lanes: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg
Lane 3: 293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 4: Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 5: Mouse lung tissue lysate at 20 µg
Lane 6: Mouse brain tissue lysate at 20 µg
All lanes: Western blot at 1/20000 dilution
Predicted band size: 85 kDa
Observed band size: 75 kDa, 85 kDa
Blocking and diluting beffer and concentration: 5% NFDM/TBST.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control at 1/200000 dilution.
The blot of Anti-Transferrin Receptor antibody [H68.4] ab269513 is applied with Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) secondary antibody at 1/10000 dilution and the blot of ab214039 is applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100000 dilution.
Lanes 1 - 9: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution
Lanes 1 - 9: Western blot - Anti-Transferrin Receptor antibody [H68.4] (Anti-Transferrin Receptor antibody [H68.4] ab269513) at 1/1000 dilution
Lane 1: Human placenta tissue lysate at 20 µg
Lane 2: Human small intestine tissue lysate at 20 µg
Lane 3: HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: LoVo (human colorectal adenocarcinoma epithelial cell)whole cell lysate at 20 µg
Lane 5: Mouse placenta tissue lysate at 20 µg
Lane 6: Mouse kidney tissue lysate at 20 µg
Lane 7: Mouse heart tissue lysate at 20 µg
Lane 8: Rat placenta tissue lysate at 20 µg
Lane 9: Rat kidney tissue lysate at 20 µg
Predicted band size: 84 kDa
Observed band size: 90 kDa
Exposure time: 26s
False colour image of Western blot: Anti-Transferrin Receptor antibody [EPR20584] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214039 was shown to bind specifically to Transferrin Receptor. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution
Lane 1: HDLM-2 cell lysate at 20 µg
Lane 2: SH-SY5Y cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 84 kDa
Observed band size: 90 kDa
Soluble transferrin receptor (sTfR) is cleaved from whole TfR at Arg 100. (PMID: 9575164, PMID: 23390091)
TfR is found as a dimer linked by disulfide bonds in cell, so sometimes we could see dimer in a few kinds of lysates. (PMID: 2507316, PMID: 3582362). This dimer was also be detected by Anti-Transferrin Receptor antibody [EPR4012] ab108985.
Blocking buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (ab214039) at 1/1000 dilution
Lane 1: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg
Lane 2: K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Exposure time: 3s
Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling Transferrin Receptor with ab214039 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
Anti-Transferrin Receptor antibody [EPR20584] ab214039 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).
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