mouse anti-human transferrin receptor (h68.4) Search Results


mouse anti-human transferrin receptor (h68.4) # 13-6890  (Sanbio Inc)
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Sanbio Inc mouse anti-human transferrin receptor (h68.4) # 13-6890
Electron microscopy images highlighting the presence of internalized EETI-II/Xfect within intracellular compartments devoid of endosomal and lysosomal markers. ( a ) Double labeling of LAMP-1 (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are LAMP-1 negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. ( b ) Double labeling of cathepsin D (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are cathepsin D negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA-GA. ( c , d ) Double labeling of <t>transferrin</t> receptor (TfR, 5 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are transferrin receptor-negative. Arrows: Transferrin receptor label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA as described in methods. ( e ) Double labeling of hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs, 10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are Hrs-negative. Arrows: Hrs label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA. ( f ) Internalized rEETI-II-Alexa488/Xfect can be found in cellular compartments including multivesicular body-like compartments (open arrowheads). Solid arrowhead: Commonly found EETI-II/Xfect positive cellular compartments of unknown nature; arrow: point of fusion between MVB-like compartment and common EETI-II/Xfect positive compartment. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. Scale bar, 200 nm.
Mouse Anti Human Transferrin Receptor (H68.4) # 13 6890, supplied by Sanbio Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Electron microscopy images highlighting the presence of internalized EETI-II/Xfect within intracellular compartments devoid of endosomal and lysosomal markers. ( a ) Double labeling of LAMP-1 (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are LAMP-1 negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. ( b ) Double labeling of cathepsin D (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are cathepsin D negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA-GA. ( c , d ) Double labeling of transferrin receptor (TfR, 5 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are transferrin receptor-negative. Arrows: Transferrin receptor label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA as described in methods. ( e ) Double labeling of hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs, 10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are Hrs-negative. Arrows: Hrs label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA. ( f ) Internalized rEETI-II-Alexa488/Xfect can be found in cellular compartments including multivesicular body-like compartments (open arrowheads). Solid arrowhead: Commonly found EETI-II/Xfect positive cellular compartments of unknown nature; arrow: point of fusion between MVB-like compartment and common EETI-II/Xfect positive compartment. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. Scale bar, 200 nm.

Journal: Scientific Reports

Article Title: Visualizing the cellular route of entry of a cystine-knot peptide with Xfect transfection reagent by electron microscopy

doi: 10.1038/s41598-019-43285-5

Figure Lengend Snippet: Electron microscopy images highlighting the presence of internalized EETI-II/Xfect within intracellular compartments devoid of endosomal and lysosomal markers. ( a ) Double labeling of LAMP-1 (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are LAMP-1 negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. ( b ) Double labeling of cathepsin D (10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are cathepsin D negative. Arrows: lysosomes. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA-GA. ( c , d ) Double labeling of transferrin receptor (TfR, 5 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are transferrin receptor-negative. Arrows: Transferrin receptor label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA as described in methods. ( e ) Double labeling of hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs, 10 nm gold particles) and Alexa488 (15 nm gold particles) shows that the majority of rEETI-II-Alexa488/Xfect positive compartments are Hrs-negative. Arrows: Hrs label. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 30 min then fixed with PFA-GA. ( f ) Internalized rEETI-II-Alexa488/Xfect can be found in cellular compartments including multivesicular body-like compartments (open arrowheads). Solid arrowhead: Commonly found EETI-II/Xfect positive cellular compartments of unknown nature; arrow: point of fusion between MVB-like compartment and common EETI-II/Xfect positive compartment. HeLa cells were treated with 5 μM rEETI-II-Alexa488/Xfect for 3 h then fixed with PFA. Scale bar, 200 nm.

Article Snippet: The antibodies applied were rabbit anti-Alexa488 (Molecular Probes, # A-11094) at 1:50; mouse anti-human LAMP-1 (CD107a) (clone H4A3, BD Pharmingen # 555801) at 1:150; goat anti-human cathepsinD (R&D Systems, # AF1014), at 1:100; rabbit anti-Hrs (M-79; Santa Cruz # sc-30221) at 1:50, and mouse anti-human transferrin receptor (H68.4; Sanbio # 13–6890) at 1:80.

Techniques: Electron Microscopy, Labeling

Internalized rEETI-II or transferrin is targeted to Xfect-induced membrane compartments in HeLa cells. HeLa cells were incubated with Xfect for indicated times then washed with PBS for three times and incubated with 0.2 mg/ml transferrin-Alexa555 ( a , b ) or 5 μM rEETI-II-Alexa488 ( c , d ) for 10 min, and washed with PBS again. Cells were fixed with 4% PFA and processed as described in methods. Fluorescence images were captured and analyzed as described in Fig. . Values were normalized to the 6 h Xfect treated samples. Mean ± SD. n = 1000 cells. Representative images from three independent experiments are shown. Scale bar, 20 µm. ( e ) EETI-II-ALexa488/Xfect is not washed away from the plasma membrane. Cells were incubated with 50 µM EETI-II-Alexa488/Xfect for 3 h, and then washed extensively with PBS before PFA fixation. The extracellular flocculent mixture of EETI-II-Alexa488 and Xfect (asterisks) sticks firmly to the cells after washing. M: microvillus. Scale bar, 200 nm.

Journal: Scientific Reports

Article Title: Visualizing the cellular route of entry of a cystine-knot peptide with Xfect transfection reagent by electron microscopy

doi: 10.1038/s41598-019-43285-5

Figure Lengend Snippet: Internalized rEETI-II or transferrin is targeted to Xfect-induced membrane compartments in HeLa cells. HeLa cells were incubated with Xfect for indicated times then washed with PBS for three times and incubated with 0.2 mg/ml transferrin-Alexa555 ( a , b ) or 5 μM rEETI-II-Alexa488 ( c , d ) for 10 min, and washed with PBS again. Cells were fixed with 4% PFA and processed as described in methods. Fluorescence images were captured and analyzed as described in Fig. . Values were normalized to the 6 h Xfect treated samples. Mean ± SD. n = 1000 cells. Representative images from three independent experiments are shown. Scale bar, 20 µm. ( e ) EETI-II-ALexa488/Xfect is not washed away from the plasma membrane. Cells were incubated with 50 µM EETI-II-Alexa488/Xfect for 3 h, and then washed extensively with PBS before PFA fixation. The extracellular flocculent mixture of EETI-II-Alexa488 and Xfect (asterisks) sticks firmly to the cells after washing. M: microvillus. Scale bar, 200 nm.

Article Snippet: The antibodies applied were rabbit anti-Alexa488 (Molecular Probes, # A-11094) at 1:50; mouse anti-human LAMP-1 (CD107a) (clone H4A3, BD Pharmingen # 555801) at 1:150; goat anti-human cathepsinD (R&D Systems, # AF1014), at 1:100; rabbit anti-Hrs (M-79; Santa Cruz # sc-30221) at 1:50, and mouse anti-human transferrin receptor (H68.4; Sanbio # 13–6890) at 1:80.

Techniques: Membrane, Incubation, Fluorescence, Clinical Proteomics