TfR (Transferrin R) Antibody (OX26)

Western Blot: TfR (Transferrin R) Antibody (OX26) [NB200-585] - Total protein from rat adrenal gland cells (PC12) and rat placenta was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/ml anti-CD71 in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence.

Immunocytochemistry/Immunofluorescence: TfR (Transferrin R) Antibody (OX26) [NB200-585] - PC-12 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti TfR/Transferrin R (OX26) NB200-585 at a 1:100 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: TfR (Transferrin R) Antibody (OX26) [NB200-585] - IHC analysis of formalin fixed paraffin-embedded (FFPE) mouse spleen using TfR (OX26) antibody at 1:50 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Plasma membrane staining was observed. Staining was performed by Histowiz.

Flow Cytometry: TfR (Transferrin R) Antibody (OX26) [NB200-585] - An intracellular stain was performed on PC12 cells with TfR (Transferrin R) [OX26] Antibody NB200-585AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: TfR (Transferrin R) Antibody (OX26) [NB200-585] - Staining of rat spleen lymphocytes with Mouse anti Rat CD71

Flow (Intracellular): TfR (Transferrin R) Antibody (OX26) [NB200-585] - PC-12 cells were stained with TfR (Transferrin R) Antibody (OX26) NB200-585 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Dylight488-conjugated anti-mouse secondary antibody.

Flow Cytometry: TfR (Transferrin R) Antibody (OX26) [NB200-585] - Analysis using the Biotin conjugate of NB200-585. Staining of mouse spleen.

Flow Cytometry: TfR (Transferrin R) Antibody (OX26) [NB200-585] - Staining of rat spleen cells with MOUSE ANTI RAT CD71: RPE.

• Reactivity: Mu, Rt
• Applications: WB, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, CyTOF-ready
• Clone: OX26
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

TfR (Transferrin R) Antibody (OX26) Summary

• Immunogen: PHA activated rat lymphocytes.
• Marker: Recycling Endosome Marker
• Specificity: Labels dividing cells of various types but also binds to a number of non-dividing normal tissues and endothelium of brain capillaries.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: TFRC
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:100-1:2000
  • Flow Cytometry 1:50-1:200
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • CyTOF-ready
  • Flow (Intracellular)
• Application Notes: Use in IHC was reported in the scientific literature (PMID: 11818402). This antibody is CyTOF ready.
• Control:

  TfR (Transferrin R) Lysate (NBP2-66234)

Reactivity Notes

Rat.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for TfR (Transferrin R) Antibody (OX26)

• CD71 antigen
• CD71
• CD71TRFR
• OX26 monoclonal antibody
• OX26 transferrin receptor
• p90
• T9
• TfR (Transferrin R)
• TfR (TransferrinR)
• TfR
• TFR1
• TFRC
• TR
• transferrin receptor (p90, CD71)
• transferrin receptor protein 1
• TRFR

Background

OX26 TfR clone is useful in studies of dividing haematopoietic and tumour cell populations, and metabolic activity. In comes in a variety of pre-conjugated fluorophores for histochemical and flow applications.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TfR (Transferrin R) Antibody (NB200-585)(14)

Publications using NB200-585

• Couty JP, Rampon C, Leveque M et al. PECAM-1 engagement counteracts ICAM-1-induced signaling in brain vascular endothelial cells. J Neurochem. [PMID: 17662049]
• Gosk S, Vermehren C, Storm G et al. Targeting anti-transferrin receptor antibody (OX26) and OX26-conjugated liposomes to brain capillary endothelial cells using in situ perfusion. J Cereb Blood Flow Metab 24(11):1193-204. 2004 Nov. [PMID: 15545912]
• Cox JH, Lamb JR, Bal V et al. The phenotypic and molecular characterization of Nb2 lymphoma cells activated with IL-2 and human growth hormone. Immunology. 1989 Jan 1 [PMID: 15493267] (FLOW, Rat)
• Moos T, Morgan EH. Restricted transport of anti-transferrin receptor antibody (OX26) through the blood-brain barrier in the rat. J Neurochem 79(1):119-29. 2001 Oct. [PMID: 11595764]
• Crook K, Hunt SV. Enrichment of early fetal-liver hemopoietic stem cells of the rat using monoclonal antibodies against the transferrin receptor, Thy-1, and MRC-OX82. Dev Immunol. [PMID: 8924759] (FLOW, Rat)
• Broadwell RD, Baker-Cairns BJ, Friden PM et al. Transcytosis of protein through the mammalian cerebral epithelium and endothelium. III. Receptor-mediated transcytosis through the blood-brain barrier of blood-borne transferrin and antibody against the transferrin receptor. Exp Neurol. 1996 Nov 1 [PMID: 8912898]
• Skarlatos S, Yoshikawa T, Pardridge WM. Transport of [125I]transferrin through the rat blood-brain barrier. Brain Res. 1995 Jun 19 [PMID: 7552351]
• Paterson DJ, Williams AF. An intermediate cell in thymocyte differentiation that expresses CD8 but not CD4 antigen. J Exp Med. 1987 Nov 1 [PMID: 3500264] (FLOW, Rat)
• de Sousa M, Dynesius-Trentham R, Mota-Garcia F et al. Activation of rat synovium by iron. Arthritis Rheum. 1988 May 1 [PMID: 3377869]
• De Sousa M, Breedvelt F, Dynesius-Trentham R et al. Iron, iron-binding proteins and immune system cells. Ann N Y Acad Sci. 1988 June 1 [PMID: 3291685]
• Opstelten D, Deenen GJ, Rozing J, Hunt SV. B lymphocyte-associated antigens on terminal deoxynucleotidyl transferase-positive cells and pre-B cells in bone marrow of the rat. J Immunol. 1986 Jul 1 [PMID: 3086459]
• Hill JM, Ruff MR, Weber RJ, Pert CB. Transferrin receptors in rat brain: neuropeptide-like pattern and relationship to iron distribution. Proc Natl Acad Sci U S A. 1985 Jul 1 [PMID: 2989832]
• Perry VH, Gordon S. Modulation of CD4 antigen on macrophages and microglia in rat brain. J Exp Med. 1987 Oct 1 [PMID: 2443599] (IHC-Fr, Rat)
• Chignola R, Colombatti M, Dell'Arciprete L et al. Distribution of endocytosed molecules to intracellular acidic environments correlates with immunotoxin activity. Int J Cancer. 1990 Dec 15 [PMID: 2123478]

FAQs for TfR (Transferrin R) Antibody (NB200-585). (Showing 1 - 2 of 2 FAQs).

1. A customer would like to use this antibody to perform an functional assay. The customer would like to know whether this antibody will block the binding of transferrin to the receptor. On the other hand, would you mind to provide the immunogenic sequence of this antibody?
   • Unfortunately, this clone has not been mapped and the sequence of the epitope recognised is currently unknown. All that is currently known is that the clone binds an extracellular region not involved in transferrin binding.
2. I was wondering if you have amino acid sequence information available for this monoclonal mouse anti-Rat TfR (Ox26) antibody.
   • The immunogen used to create this product is PHA activated rat lymphocytes (SwissProt: Q99376). We have not mapped the epitope for the product.

Bioinformatics

• Gene Symbol: TFRC
• Entrez:
  • 7037(Human)
  • 22042(Mouse)
  • 64678(Rat)
• Uniprot:
  • Q99376(Rat)