Cells were lysed with NP-40 lysis buffer (25 mM tris (pH 7.4), 1% Triton X-100, 1% SDS, 1% sodium deoxy-cholate, 150 mM NaCl, aprotinin (2 μg/ml), 1 mM phenylmethylsulfonyl fluoride) containing complete protease inhibitor cocktail (Roche Diagnostics). Samples were separated by SDS–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes before being incubated with primary and secondary antibodies. Bands were detected using chemiluminescence (Thermo Scientific). Membranes were probed with antibodies to β-actin (Sigma, Cat# A3854 ), ferroportin (Novus Biologicals, cat# NBP121502 ), transferrin receptor1 (Invitrogen, cat# 13-6890), IRP2 (Santa Cruz, cat# sc-33682), ferritin H (17 (link)), p-STAT3 (cell Signaling, cat# 9131S) and STAT3 (cell Signaling, cat#4904S ). Western blots were quantified using ImageJ software.
Transferrin receptor1
Manufactured by Thermo Fisher Scientific
Transferrin receptor1 is a protein that is responsible for the cellular uptake of iron by facilitating the internalization of iron-bound transferrin. It is a key regulator of iron homeostasis in the body.
Automatically generated - may contain errors
2 protocols using transferrin receptor1
1
Western Blot Analysis of Iron Homeostasis
2
Western Blot Analysis of Iron Homeostasis
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Cells were lysed with NP-40 lysis buffer (25 mM tris (pH 7.4), 1% Triton X-100, 1% SDS, 1% sodium deoxy-cholate, 150 mM NaCl, aprotinin (2 μg/ml), 1 mM phenylmethylsulfonyl fluoride) containing complete protease inhibitor cocktail (Roche Diagnostics). Samples were separated by SDS–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes before being incubated with primary and secondary antibodies. Bands were detected using chemiluminescence (Thermo Scientific). Membranes were probed with antibodies to β-actin (Sigma, Cat# A3854 ), ferroportin (Novus Biologicals, cat# NBP121502 ), transferrin receptor1 (Invitrogen, cat# 13-6890), IRP2 (Santa Cruz, cat# sc-33682), ferritin H (17 (link)), p-STAT3 (cell Signaling, cat# 9131S) and STAT3 (cell Signaling, cat#4904S ). Western blots were quantified using ImageJ software.
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