- MA5-11441 - Invitrogen Antibodies TFRC antibody

Submitted references A novel manganese complex selectively induces malignant glioma cell death by targeting mitochondria.

Geng J, Li J, Huang T, Zhao K, Chen Q, Guo W, Gao J
Molecular medicine reports 2016 Sep;14(3):1970-8

Artemisinin derivatives induce iron-dependent cell death (ferroptosis) in tumor cells.

Ooko E, Saeed ME, Kadioglu O, Sarvi S, Colak M, Elmasaoudi K, Janah R, Greten HJ, Efferth T
Phytomedicine : international journal of phytotherapy and phytopharmacology 2015 Oct 15;22(11):1045-54

Spatial and temporal organization of tick-borne encephalitis flavivirus replicated RNA in living cells.

Miorin L, Maiuri P, Hoenninger VM, Mandl CW, Marcello A
Virology 2008 Sep 15;379(1):64-77

Isolation and characterization of putative epidermal stem cells derived from Cashmere goat fetus.

Islam MS, Zhou H
European journal of dermatology : EJD 2007 Jul-Aug;17(4):302-8

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of CD71 was performed by loading 25 µg of Hela (Lane 1), MCF-7 (Lane 2), and NIH-3T3 (Lane 3) cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4ºC overnight. The membrane was probed with a CD71 monoclonal antibody (Product # MA5-11441) at a dilution of 1:200 (Hela) and 1:100 (MCF-7 and NIH-3T3) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 95 kDa in Hela cell lysates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Transferrin Receptor was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR944258_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Transferrin Receptor was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) andHeLa Transferrin Receptor KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Transferrin Receptor Monoclonal Antibody (10F11) (Product # MA5-11441, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Transferrin Receptor.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of CD71 showing positive staining in the cytoplasm and membrane of paraffin-treated Human placenta tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CD71 monoclonal antibody (Product # MA5-11441) diluted by 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of CD71 in Jurkat cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CD71 monoclonal antibody (Product # MA5-11441) at a dilution of 1:2 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CD71 in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CD71 monoclonal antibody (Product # MA5-11441) at a dilution of 1:2 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Adpa-manganese (Mn) exhibited selective inhibition on glioma cell proliferation. (A) Structure of Adpa-Mn. (B and C) U251 and C6 cells were exposed to various concentrations of Adpa-Mn, and cell proliferation was determined by 3-[4,5-dimehyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bro mide (MTT) assay every 12 h. (D) C6 cells and rat astrocyte cells (AST) were exposed to various concentrations of Adpa-Mn for 24 h, and cell proliferation was determined by MTT assay. (E) Images of cellular morphological alterations were captured (scale bar, 10 u m). (F) C6 cells and AST cells were exposed to various concentrations of cisplatin for 24 h, and cell proliferation was determined by MTT assay. (G) Expression of divalent metal transporter 1 (DMT-1) and transferrin receptor (TfR) in U251 cells following ferric citrate (100 u M) or desferrioxamine (DFO; 10 u M) pretreatment for 24 h, as determined by western blotting. (H) U251 cells were pretreated with ferric citrate (100 u M) or DFO (10 u M) for 24 h, and were then treated with Adpa-Mn for 24 h. Cell proliferation was determined by MTT assay. Data are presented as the mean +- standard deviation of three independent experiments. * P

MA5-11441

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