Anti tfr

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-TfR is a lab equipment product that targets the transferrin receptor (TfR) protein. It is used for research purposes in various applications related to cellular and molecular biology.

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Lab products found in correlation

Saf32, by Cayman Chemical (2 mentions) Anti β actin mab1501, by Merck Group (2 mentions) Na934v, by GE Healthcare (2 mentions) Anti gapdh gt239, by GeneTex (2 mentions) Anti ferritin, by Merck Group (2 mentions) Anti tf gtx2123, by GeneTex (2 mentions) Anti ceruloplasmin, by Agilent Technologies (2 mentions) Anti fpn nbp1 21502, by Novus Biologicals (2 mentions) Hoechst, by Thermo Fisher Scientific (2 mentions) Fluoromount g, by Southern Biotech (2 mentions) Pngase f, by New England Biolabs (2 mentions) Dcs 6, by BD (1 mentions) Anti ralb, by Cell Signaling Technology (1 mentions) Immobilon western, by Merck Group (1 mentions) Anti ha, by Roche (1 mentions) Anti tubulin, by Merck Group (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Subcellular protein fractionation kit for cultured cells, by Thermo Fisher Scientific (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Anti pcna, by Abcam (1 mentions)

Close spelling variants of this product

Anti-Transferrin receptor (TfR) (2 citations) Mouse anti-TfR (2 citations)

7 protocols using anti tfr

Subcellular Protein Fractionation and Immunoblotting

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Protein fractionation was performed with the Subcellular Protein Fractionation kit for cultured cells (Thermo Scientific-Pierce; 78840). Soluble fraction corresponds to a mixture of cytosolic and nuclear soluble fractions described in the supplier's instructions.
For immunoblot, protein samples were resolved by SDS-PAGE, transferred to PVDF membranes (Millipore), and incubated with primary antibodies anti-Ccnd1 (monoclonal DCS-6, BD Pharmigen), anti-Cdk4 (polyclonal C-22, sc-260), anti-HA (rat monoclonal 3F10, Roche), anti-TfR (monoclonal H68.4, Invitrogen), anti-RalB (rabbit polyclonal, Cell Signaling), anti-PCNA (monoclonal PC10, Abcam) and anti-tubulin (monoclonal B-5-1-2 Sigma). Appropriate peroxidase-linked secondary antibodies (GE Healthcare UK Ltd) were detected using the chemiluminescent HRP substrate Immobilon Western (Millipore). Chemiluminescence was recorded with a ChemiDoc-MP imaging system (BioRad).

Fusté N.P., Castelblanco E., Felip I., Santacana M., Fernández-Hernández R., Gatius S., Pedraza N., Pallarés J., Cemeli T., Valls J., Tarres M., Ferrezuelo F., Dolcet X., Matias-Guiu X, & Garí E. (2016). Characterization of cytoplasmic cyclin D1 as a marker of invasiveness in cancer. Oncotarget, 7(19), 26979-26991.

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Immunofluorescence Labeling of Endocytic Organelles

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Cells were enzymatically labeled, fixed in 2% paraformaldehyde, 7% sucrose, in PBS. Free aldehydic groups from paraformaldehyde were covered incubating with 50 mM NH₄Cl for 10 min. Buffer containing 0.25% Saponin, 5% normal goat serum, and 2% serum bovine albumine in PBS was used for permeabilization and blocking; incubation with antibodies was performed in the presence of 0.25% Saponin. The following primary antibodies were used: mouse antiearly endosome antigen 1 (anti-EEA1) 1:100 (610456, BD Biosciences, San Jose, CA, USA); rabbit antilysosomal-associated membrane protein 1 (anti-LAMP1) 1:500 (ab24170; abcam, Cambridge, UK); rabbit anti-Rab4 1:100 (sc-312, Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit anti-Rab5 1:100 (sc-28570, Santa Cruz Biotechnology); rabbit anti-Rab7 1:100 (sc-10767, Santa Cruz Biotechnology); rabbit anti-Rab11 10 µg/ml (71-5300, Invitrogen); mouse antivesicle-associated membrane protein 2 (anti-VAMP2) 1:500 (Synaptic Systems, Gottingen, Germany); rabbit anti-VAMP3 1:200 (ThermoFisher Scientific); rabbit anti-Syntaxin-6 1:100 (Synaptic Systems); and mouse antitransferrin receptor (anti-TfR) 1:200 (Invitrogen). The secondary antibodies used were: goat anti-Mouse Alexa Fluor^® 405 conjugate 1:250 (Invitrogen); goat anti-Rabbit DyLight 405 conjugate 1:500 (ThermoFisher Scientific). Immunofluorescence microscopy was performed as described before.

Mazzotti C., Gagliostro V., Bosisio D., Del Prete A., Tiberio L., Thelen M, & Sozzani S. (2017). The Atypical Receptor CCRL2 (C-C Chemokine Receptor-Like 2) Does Not Act As a Decoy Receptor in Endothelial Cells. Frontiers in Immunology, 8, 1233.

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Antibody Panel for HIV Protein Analysis

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We listed all antibodies used in this study, anti-Gag (made by ourselves); anti HIV-1 Env gp120 (Abcam); anti-CCDC8 (Cat#: H00083987-B01P, Abnova); anti-Caveolin-1 (Santa Cruz Biotechnology); anti-TFR (Invitrogen); anti-His (Invitrogen), anti-Flag (Invitrogen), anti-HA (Invitrogen), anti-V5 (Invitrogen), anti-CD63 (Abcam), anti-26S proteasome marker PSMD8 (Cat #: H00005714-B01, Abnova) and secondary antibody anti-mouse (Invitrogen), anti-rabbit (Invitrogen).

Wei M., Zhao X., Liu M., Huang Z., Xiao Y., Niu M., Shao Y, & Kleiman L. (2015). Inhibition of HIV-1 assembly by coiled-coil domain containing protein 8 in human cells. Scientific Reports, 5, 14724.

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Western Blotting of Caveolin and Transferrin Receptor

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Aliquots (25 μl) were taken from each fraction, mixed with Laemmli sample buffer (Bio-Rad 161-0737) containing 5% 2-mercaptoethanol, separated by SDS-PAGE on a 4–15% polyacrylamide gel, and transferred to nitrocellulose membranes (Pierce Biotechnology, Rockford, IL) for Western blotting. Membranes were probed with a primary antibody to caveolin (1:1000 anti-caveolin, BD Transduction Laboratories, 610059) or transferrin receptor (1:1000 anti-TfR, Invitrogen, 13-6800). Secondary antibodies included goat anti-mouse or anti-rabbit conjugated to horseradish peroxidase (1:10000, Thermo Scientific Pierce, 31432 or 31460). Reactions were visualized with SuperSignal West Femto chemiluminescent substrate (Thermo Fisher Pierce, 34095) and imaged on an Alpha Innotech FluorChem SP Imaging System (ProteinSimple).

Thomas P.V., Cheng A.L., Colby C.C., Liu L., Patel C.K., Josephs L, & Duncan R.K. (2014). LOCALIZATION AND PROTEOMIC CHARACTERIZATION OF CHOLESTEROL-RICH MEMBRANE MICRODOMAINS IN THE INNER EAR. Journal of proteomics, 103, 178-193.

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Antibodies for Protein Analysis

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The following antibodies were used in this study: Anti-PrP antibodies 3F4 and 8H4 (Signet Laboratories, Dedham, MA), and SAF32 (189720, Cayman chemicals, USA), anti-ferritin (F5012, Sigma Aldrich, USA), anti-TfR (136800, Invitrogen, USA), anti-Tf (GTX2123, GeneTEX, USA), anti-ceruloplasmin (Dako, USA), Mouse IgG (5415, Cell signaling Technology, USA), anti-Fpn (NBP1-21502, Novus biological, USA), anti β-actin (MAB1501, Milipore, USA) and anti GAPDH (GT239, GeneTEX). HRP-conjugated anti-mouse, NA931V and anti-rabbit, NA934V (GE Healthcare, UK). Alexa Fluor-conjugated secondary antibodies (molecular probes), PNGase F (P0704S, NEB, USA). NAP (Non-animal protein) blocking solution (786-190T, G-Biosciences, USA), Hoechst (#33342, Invitrogen, USA), Fluoromount-G (Southern Biotech, USA).

Ashok A., Karmakar S., Chandel R., Ravikumar R., Dalai S., Kong Q, & Singh N. (2018). Prion protein modulates iron transport in the anterior segment: Implications for ocular iron homeostasis and prion transmission. Experimental eye research, 175, 1-13.

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Antibodies for Protein Analysis

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Immunofluorescent Localization of Organelle Markers

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Cells were fixed in 4% paraformaldehyde in PBS and permeabilized/blocked with 0.1% saponin/5% goat serum in PBS (a methanol permeabilization step was included for anti-PDI labelling). Antibody incubations were performed in PBS/0.01% saponin/5% goat serum. The primary antibodies used were anti-RFP (rat monoclonal 5F8; antibodies-online.com), anti-mCherry (mouse monoclonal 1C51; Novus Biologicals), anti-Lamp1 (rat monoclonal 1D4B; DSHB), anti-TfR (mouse monoclonal H68.4; Invitrogen), anti-EEA1 (rabbit monoclonal, C45B10; Cell Signalling Technology), and anti-PDI (rabbit monoclonal, C81H6; Cell Signalling Technology). The secondary antibodies used were derived from goat serum, cross-absorbed, and conjugates of Alexa 488 (for organelle markers) or Alexa 546 (for mCherry) (Invitrogen). Cells were viewed on a Zeiss 510 META confocal microscope, in multitrack mode, using the following excitation/emission parameters (nm): Alexa 488 (488/505–530) and Alexa 546 (543/>560).

Ruas M., Davis L.C., Chen C.C., Morgan A.J., Chuang K.T., Walseth T.F., Grimm C., Garnham C., Powell T., Platt N., Platt F.M., Biel M., Wahl-Schott C., Parrington J, & Galione A. (2015). Expression of Ca2+-permeable two-pore channels rescues NAADP signalling in TPC-deficient cells. The EMBO Journal, 34(13), 1743-1758.

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