Journal:

Article Title: Cloning and Characterization of a Calcium-binding, Histamine-releasing Protein from Schistosoma mansoni *

doi: 10.1074/jbc.M204114200

Figure Lengend Snippet: A, ~20 μg of protein extracted from uninduced (Uind) and induced (Ind) cultures of Escherichia coli containing SmTCTP expression construct were separated in a 12% SDS-PAGE. rSmTCTP was then purified from the cultures using a nickel-nitrilotriacetic acid chromatography column, and ~1 μg of the protein was separated in a 12% SDS-PAGE (rSmTCTP). B, affinity-purified rSmTCTP probed with sera collected from C57BL/6 mice on day 0 or week 2, 3, 8, or 9 after infection with S. mansoni. Purified rSmTCTP probed with a mouse anti-SmTCTP serum served as a positive control.

Article Snippet: Briefly, purified rSmTCTP transblotted onto nitrocellulose membranes was incubated with 20 μ Ci/ml 45 CaCl 2 (ICN Biomedicals, Costa Mesa, CA) for 10 min, washed with distilled water for 5 min, air-dried, and exposed to x-ray film for 10 min.

Techniques: Expressing, Construct, SDS Page, Purification, Chromatography, Affinity Purification, Infection, Positive Control

Journal:

Article Title: Cloning and Characterization of a Calcium-binding, Histamine-releasing Protein from Schistosoma mansoni *

doi: 10.1074/jbc.M204114200

Figure Lengend Snippet: About 1 μg of purified rSmTCTP was transferred to nitrocellulose membrane and probed with radioactive 45CaCl2 as described under “Experimental Procedures.” Bovine muscle tropomyosin was a positive control, and bovine serum albumin (BSA) was used as a negative control. The data shown are representative of two similar experiments.

Article Snippet: Briefly, purified rSmTCTP transblotted onto nitrocellulose membranes was incubated with 20 μ Ci/ml 45 CaCl 2 (ICN Biomedicals, Costa Mesa, CA) for 10 min, washed with distilled water for 5 min, air-dried, and exposed to x-ray film for 10 min.

Techniques: Purification, Positive Control, Negative Control

Journal:

Article Title: Cloning and Characterization of a Calcium-binding, Histamine-releasing Protein from Schistosoma mansoni *

doi: 10.1074/jbc.M204114200

Figure Lengend Snippet: About 1 × 105 RBL-2H3 cells were incubated with 10, 1.25, or 0.3 μg/ml rSmTCTP for 30 min at 37 °C; the amount of histamine released into the culture supernatant was measured using a kit. As a positive control, 48/80 was added to the wells at a concentration of 0.3 μg/ml. Recombinant SmGBF was used as a negative control. The data shown are the mean ± S.D. of five different experiments. All three concentrations of rSmTCTP released significantly higher amounts of histamine than found in negative controls or upon spontaneous release (p < 0.01).

Article Snippet: Briefly, purified rSmTCTP transblotted onto nitrocellulose membranes was incubated with 20 μ Ci/ml 45 CaCl 2 (ICN Biomedicals, Costa Mesa, CA) for 10 min, washed with distilled water for 5 min, air-dried, and exposed to x-ray film for 10 min.

Techniques: Incubation, Positive Control, Concentration Assay, Recombinant, Negative Control

Journal:

Article Title: Cloning and Characterization of a Calcium-binding, Histamine-releasing Protein from Schistosoma mansoni *

doi: 10.1074/jbc.M204114200

Figure Lengend Snippet: C57BL/6 mice were sensitized 1 week prior to the injection of rSmTCTP with ovalbumin. Peritoneal cells were collected 24 h after injection with 5 μg of endotoxin-free rSmTCTP, and a differential count was made on a cytospin smear preparation stained with Giemsa. Control animals received 100 μl of sterile saline. The data presented are from one of three identical experiments with similar results. Values shown are the mean (±S.E.) from five animals/group. *, significant at p < 0.01.

Article Snippet: Briefly, purified rSmTCTP transblotted onto nitrocellulose membranes was incubated with 20 μ Ci/ml 45 CaCl 2 (ICN Biomedicals, Costa Mesa, CA) for 10 min, washed with distilled water for 5 min, air-dried, and exposed to x-ray film for 10 min.

Techniques: Injection, Staining