Journal: Bioconjugate Chemistry

Article Title: Enhancing Reactivity for Bioorthogonal Pretargeting by Unmasking Antibody-Conjugated trans-Cyclooctenes

doi: 10.1021/bc500605g

Figure Lengend Snippet: Figure 5. Translation of dual orthogonal coupling to different antibodies. (A) Anti-TfR and EpCAM antibodies had low TCO reactivity (12−15%) after direct amine-conjugation, but the PEG24− TCO increased reactive TCO density 3- to 4-fold. *MALDI-TOF analysis of the TCO−PEG24-modified anti-EpCAM antibody was limited by difficulties with desorption. (B) Flow cytometry results for TCO- and TCO−PEG24-modified anti-EpCAM and TfR antibodies on A431 cells after reaction with 10 nM tetrazine−QDs for 30 min, showing 5-fold signal enhancements for the PEG24−TCO. (C) Confocal images of A431 cells labeled with (i, iv) anti-EpCAM, (ii, v) anti-TfR, or (iii, vi) nonbinding control antibodies. The antibodies were modified with (i−iii) TCO or (iv−vi) PEG24−TCO at the highest densities and reacted with tetrazine−QD at 10 nM for 30 min. Scale bar: 20 μm. Error bars represent the standard error of at least three independent experiments.

Article Snippet: Monoclonal mouse antibodies specific for human EpCAM (IgG2b, clone 158206) and transferrin receptor (TfR, IgG1, clone 29806) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Conjugation Assay, Flow Cytometry, Labeling, Control

Journal: The American Journal of Pathology

Article Title: Regulation of Apo2L/Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Induced Apoptosis in Thyroid Carcinoma Cells

doi: 10.1016/s0002-9440(10)64220-4

Figure Lengend Snippet: Figure 10. A: SW579 cells were sensitized by a 48-hour pretreatment with IFN- (500 IU/ml) and TNF- (50 ng/ml) to an additional 18-hour incubation with a low, subtoxic concentration of LZ-TRAIL (20 ng/ml). B: TRAIL- resistant SW579-TR cells were sensitized by a 48-hour pretreatment with IFN- (500 IU/ml) and TNF- (50 ng/ml) to an additional 18-hour incubation with LZ-TRAIL (300 ng/ml).

Article Snippet: Recombinant human TRAIL was obtained from Immunex Corporation (Seattle, WA), in a leucine zipper (LZ) form that promotes and stabilizes the formation of trimers, as previously reported;13 for comparison, several experiments were repeated using the recombinant Apo2L form from Genentech Inc. (South San Francisco, CA).20 The goat polyclonal antibodies for DR4, DR5, DcR1, and mouse monoclonal antibody for tubulin were from Santa Cruz Biotechnologies (Santa Cruz, CA); the anti-DcR2 rabbit polyclonal antibody was from Imgenex (San Diego, CA); cycloheximide, geldanamycin, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was from Sigma Chemical Co. (St Louis, MO); IGF-1, bFGF, EGF, IFN- , and TNF- were from R&D Systems (Minneapolis, MN; bisindolylmaleimide (BIM) III and wortmannin were from Calbiochem (La Jolla, CA); rabbit polyclonal antibody for caspase-10 was from Research Diagnostics Inc. (Flanders, NJ); the IGF-1 receptor neutralizing antibody aIR3 was from Oncogene Research (Cambridge, MA); and the enhanced chemiluminescence (ECL) kit, which includes the peroxidase-labeled anti-mouse and anti-rabbit secondary antibodies, was from Amersham (Arlington Heights, IL).

Techniques: Incubation, Concentration Assay

Journal: The American Journal of Pathology

Article Title: Regulation of Apo2L/Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Induced Apoptosis in Thyroid Carcinoma Cells

doi: 10.1016/s0002-9440(10)64220-4

Figure Lengend Snippet: Figure 11. A and B: Flow cytometric analysis of TRAIL receptors DR4 (A) and DR5 (B) after a 48-hour treatment with or without IFN- (500 IU/ml) or TNF- (50 ng/ml) in SW579 cells. Control antibody staining is also shown. C: Evaluation of the protein levels of caspase-8, caspase-10, caspase-3, FLIP, and TRAIL in SW579 cells after a 48-hour treatment with or without IFN- (500 IU/ml) or TNF- (50 ng/ml). IFN- (500 IU/ml) up-regulated caspase-8 and TNF- (50 ng/ml) and up-regulated caspases-10 and -3. Additionally, TNF- induced the expression of TRAIL itself.

Article Snippet: Recombinant human TRAIL was obtained from Immunex Corporation (Seattle, WA), in a leucine zipper (LZ) form that promotes and stabilizes the formation of trimers, as previously reported;13 for comparison, several experiments were repeated using the recombinant Apo2L form from Genentech Inc. (South San Francisco, CA).20 The goat polyclonal antibodies for DR4, DR5, DcR1, and mouse monoclonal antibody for tubulin were from Santa Cruz Biotechnologies (Santa Cruz, CA); the anti-DcR2 rabbit polyclonal antibody was from Imgenex (San Diego, CA); cycloheximide, geldanamycin, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was from Sigma Chemical Co. (St Louis, MO); IGF-1, bFGF, EGF, IFN- , and TNF- were from R&D Systems (Minneapolis, MN; bisindolylmaleimide (BIM) III and wortmannin were from Calbiochem (La Jolla, CA); rabbit polyclonal antibody for caspase-10 was from Research Diagnostics Inc. (Flanders, NJ); the IGF-1 receptor neutralizing antibody aIR3 was from Oncogene Research (Cambridge, MA); and the enhanced chemiluminescence (ECL) kit, which includes the peroxidase-labeled anti-mouse and anti-rabbit secondary antibodies, was from Amersham (Arlington Heights, IL).

Techniques: Control, Staining, Expressing

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: Anti-Cancer Activity of Doxorubicin-Loaded Liposomes Co-Modified with Transferrin and Folic Acid

doi: 10.1016/j.ejpb.2016.05.023

Figure Lengend Snippet: Characterization of Liposomes

Article Snippet: Materials, Cell Culture and Animals FITC-labeled mouse monoclonal anti-transferrin receptor antibody (ab47095) was purchased from Abcam (San Francisco, CA); goat anti-folate receptor alpha primary antibody (sc-16386), FITC-labeled secondary donkey anti-goat antibody (sc-2024) and FITC-labeled normal mouse IgG (sc-2855) (as negative control) were purchased from Santa Cruz Biotechnology (Dallas, TX); nitrophenylcarbonyl-PEG 3400 -nitrophenylcarbonyl (NPC-PEG-NPC) and amino-PEG 3400 -DSPE were from Laysan Bio (Arab, AL); eggphosphatidylcholine (ePC), cholesterol, cholesteryl hemisuccinate (CHEMS), 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE), 1,2-distearoyl- sn -glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG), (lissamine rhodamine) DPPE (rhodamine) were from Avanti (Alabaster, AL); Dragendorff’s reagent, Molybdenum blue, triethylamine (TEA), Sepharose CL-4B (40–165 μm), transferrin, folic acid, dicyclohexylcarbodiimide (DCC) and Ninhydrin were from Sigma-Aldrich (St.Louis, MO); microBCA ™ protein assay kit, Whatman nucleopore polycarbonate membranes 19 mm at 0.2μ, 0.1μ and 0.05μ pore sizes, bovine serum albumin (BSA) and agarose were from Fisher/Thermo Scientific (Waltham, MA); Spectra/Por pre-wetted 300,000 MWCO dialysis membranes and Spectra/Por 12–14 kDa MWCO dry membranes were from Spectrum Labs (Rancho Dominguez, CA); CellTiter-Blue ® cell viability assay was from Promega Corporation (Madison, WI); doxorubicin hydrochloride was from LC Labs (Woburn, MA) and Lancrix Chemicals (Shanghai, China).

Techniques:

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: Anti-Cancer Activity of Doxorubicin-Loaded Liposomes Co-Modified with Transferrin and Folic Acid

doi: 10.1016/j.ejpb.2016.05.023

Figure Lengend Snippet: Characterization of (A) transferrin and (B) folate receptor expression.

Article Snippet: Materials, Cell Culture and Animals FITC-labeled mouse monoclonal anti-transferrin receptor antibody (ab47095) was purchased from Abcam (San Francisco, CA); goat anti-folate receptor alpha primary antibody (sc-16386), FITC-labeled secondary donkey anti-goat antibody (sc-2024) and FITC-labeled normal mouse IgG (sc-2855) (as negative control) were purchased from Santa Cruz Biotechnology (Dallas, TX); nitrophenylcarbonyl-PEG 3400 -nitrophenylcarbonyl (NPC-PEG-NPC) and amino-PEG 3400 -DSPE were from Laysan Bio (Arab, AL); eggphosphatidylcholine (ePC), cholesterol, cholesteryl hemisuccinate (CHEMS), 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE), 1,2-distearoyl- sn -glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG), (lissamine rhodamine) DPPE (rhodamine) were from Avanti (Alabaster, AL); Dragendorff’s reagent, Molybdenum blue, triethylamine (TEA), Sepharose CL-4B (40–165 μm), transferrin, folic acid, dicyclohexylcarbodiimide (DCC) and Ninhydrin were from Sigma-Aldrich (St.Louis, MO); microBCA ™ protein assay kit, Whatman nucleopore polycarbonate membranes 19 mm at 0.2μ, 0.1μ and 0.05μ pore sizes, bovine serum albumin (BSA) and agarose were from Fisher/Thermo Scientific (Waltham, MA); Spectra/Por pre-wetted 300,000 MWCO dialysis membranes and Spectra/Por 12–14 kDa MWCO dry membranes were from Spectrum Labs (Rancho Dominguez, CA); CellTiter-Blue ® cell viability assay was from Promega Corporation (Madison, WI); doxorubicin hydrochloride was from LC Labs (Woburn, MA) and Lancrix Chemicals (Shanghai, China).

Techniques: Expressing

Journal: Journal of Biological Chemistry

Article Title: Post-translational Modifications of Three Members of the Human MAP1LC3 Family and Detection of a Novel Type of Modification for MAP1LC3B

doi: 10.1074/jbc.m303800200

Figure Lengend Snippet: FIG. 6. Subcellular distribution of rat MAP1LC3 and three human homologues. HeLa cells (T) transfected with myc-LC3A (A), myc-LC3B (B), myc-LC3 (C), and myc-rat LC3 (D) were homogenized and fractionated into supernatant (S) and pellet (P) by ultracentrifugation. These fractions were analyzed by Western blotting using antibodies against Myc tag, transferrin receptor (as a membrane protein marker), and aldolase (as a cytosolic marker). The asterisk indicates that the band only existed in the HeLa cells transfected with Myc-LC3C.

Article Snippet: Antibodies against human transferrin receptor and human aldolase were purchased from Santa Cruz Biotechnology, Inc.

Techniques: Transfection, Western Blot, Membrane, Marker