Protein extracts were prepared with cytoplasmic lysis buffer (25mM Tris-HCl [pH7.4], 40mM KCl, and 1% Triton X-100) supplemented with 1 mg/ml aprotinin and 1 mg/ml leupeptin (all obtained from Sigma). Twenty micrograms of total protein was run on 10%−15% SDS-polyacrylamide gels, and Western blotting was performed exactly as described (Theurl et al., 2006 (link)) with a mouse anti-human TfR1 antibody (1:1000; Invitrogen), rabbit anti-human Ferritin (1:500; Sigma), rabbit anti-Fpn (1:400; self designed; Eurogentic), rabbit anti-HO-1 (1:1000; Enzo), rabbit anti-iNOS (1:1000; BD), rabbit anti-NFκB p65 or rabbit anti-phosphor NFκB p65 (both 1:1000; Cell Signalling). Blotting with either rabbit anti-TBP (1:1000; Cell Signalling) or rabbit anti-Actin antibody (1:1000; Sigma-Aldrich) was performed as a loading control.
Mouse anti human tfr1 antibody
Manufactured by Thermo Fisher Scientific
The Mouse anti-human TfR1 antibody is a laboratory reagent used for the detection and analysis of the human Transferrin Receptor 1 (TfR1) protein. TfR1 is a cell surface receptor involved in the cellular uptake of iron through the binding and internalization of transferrin. This antibody can be used in various immunoassay techniques to identify and quantify the expression of TfR1 in research samples.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti human ferritin, by Merck Group (2 mentions)
Aprotinin, by Merck Group (2 mentions)
Leupeptin, by Merck Group (2 mentions)
Rabbit anti actin antibody, by Merck Group (1 mentions)
Rabbit anti nf κb p65, by Cell Signaling Technology (1 mentions)
Rabbit anti tbp, by Cell Signaling Technology (1 mentions)
Rabbit anti phosphor nfκb p65, by Cell Signaling Technology (1 mentions)
Rabbit anti inos, by BD (1 mentions)
Rabbit anti ho 1, by Enzo Life Sciences (1 mentions)
Rabbit anti β actin antibody, by Merck Group (1 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
2 protocols using mouse anti human tfr1 antibody
1
Western Blot Analysis of Iron Metabolism Proteins
2
Quantitative Western Blot Analysis of TfR1, Ferritin, and p4E-BP1
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Protein extracts were prepared with cytoplasmic lysis buffer (25 mM Tris–HCl [pH 7.4], 40 mM KCl, and 1% Triton X-100) supplemented with 1 mg/ml aprotinin and 1 mg/ml leupeptin (all obtained from Sigma). 20 μg of total protein were run on 10–15% SDS-polyacrylamide gels, and western blotting was performed using the PVDF transfer membrane as previously described [48 (link)]. We used either a mouse anti-human TfR1 antibody (1:1000; Invitrogen, rabbit anti-human ferritin (1:500, Sigma) or rabbit anti-mouse p4E-BP1 (1:1000; Cellsignal) antibodies. Blotting with rabbit anti-β-Actin antibody (1:500; Sigma-Aldrich) was performed as a loading control. The chemiluminescence signal was detected with a ChemiDoc Imaging system (BioRad) and densitometric scanning of blots was carried out for quantitative comparisons.
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