Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)

Rabbit Recombinant Monoclonal Transferrin Receptor antibody. Carrier free. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples.

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Images

Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (AB232376), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	IHC-P
Human	Tested	Tested	Expected	Tested
Mouse	Expected	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information TFRC

Function

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.

Alternative names

CD71, Transferrin receptor protein 1, TR, TfR, TfR1, Trfr, T9, p90, TFRC

Recommended products

Rabbit Recombinant Monoclonal Transferrin Receptor antibody. Carrier free. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR20584
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab232376 is the carrier-free version of Anti-Transferrin Receptor antibody [EPR20584] ab214039.

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

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13 product images

Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
Transferrin Receptor was immunoprecipitated from 0.35 mg of K562 (human chronic myelogenous leukemia cell line from bone marrow) lysate with Anti-Transferrin Receptor antibody [EPR20584] ab214039 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Transferrin Receptor antibody [EPR20584] ab214039 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: K562 whole cell lysate 10 μg (Input).
Lane 2: Anti-Transferrin Receptor antibody [EPR20584] ab214039 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Transferrin Receptor antibody [EPR20584] ab214039 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
All lanes: Immunoprecipitation - Anti-Transferrin Receptor antibody [EPR20584] (Anti-Transferrin Receptor antibody [EPR20584] ab214039)
Predicted band size: 84 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Heat mediated antigen retrieval was performed 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] - BSA and Azide free (ab232376)
Immunohistochemical analysis of paraffin-embedded human esophageal cancer tissue labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Heat mediated antigen retrieval was performed 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] - BSA and Azide free (ab232376)
Immunohistochemical analysis of paraffin-embedded human esophagus tissue labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Heat mediated antigen retrieval was performed 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] - BSA and Azide free (ab232376)
Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
Immunofluorescent analysis of 100% methanol-fixed K562 (human chronic myelogenous leukemia cell line from bone marrow) cells labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
Immunofluorescent analysis of 100% methanol-fixed RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
All lanes: Western blot - Anti-Transferrin Receptor antibody [EPR20584] (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
Secondary
All lanes: Western blot at 1/20000 dilution
Predicted band size: 85 kDa
Observed band size: 75 kDa, 85 kDa
Western blot - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039).
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 Anti-Transferrin Receptor antibody [EPR20584] 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] (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
Western blot - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Transferrin Receptor antibody [EPR20584] ab214039). 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, Anti-Transferrin Receptor antibody [EPR20584] 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] (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
Western blot - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
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] (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
Secondary
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody [EPR20584] - BSA and Azide free (ab232376)
This data was developed using Anti-Transferrin Receptor antibody [EPR20584] ab214039, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling Transferrin Receptor with Anti-Transferrin Receptor antibody [EPR20584] 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] 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).