Journal:

Article Title: Identification of Polymorphonuclear Leukocyte and HL-60 Cell Receptors for Adhesins of Streptococcus gordonii and Actinomyces naeslundii

doi:

Figure Lengend Snippet: Lectin blotting of extracts of HL-60 cells and PMNs. Cellular extracts (5 μg of protein per lane) from HL-60 cells prior to (⊘) and during DMSO-induced differentiation as well as from PMNs were separated by SDS-PAGE. Nitrocellulose transfers were untreated or treated with sialidase prior to incubation with biotinylated LFA, PNA, or ConA. Bound lectins were detected with avidin-d-alkaline phosphatase. Lanes containing either fetuin (F), asialofetuin (AF), or thyroglobulin (TG) (5 μg per lane) were included as controls. Sizes of molecular-mass markers are indicated to the left in kilodaltons.

Article Snippet: Transfers were subsequently incubated for 1 h at room temperature with biotinylated Limax flavus agglutinin (LFA), biotinylated peanut agglutinin (PNA), or biotinylated Canavalia ensiformis agglutinin (ConA) (EY Laboratories, Inc., San Mateo, Calif.) at concentrations of 5 μg of LFA or PNA per ml or 50 μg of ConA per ml in blocking buffer.

Techniques: SDS Page, Incubation, Avidin-Biotin Assay

Journal:

Article Title: Identification of Polymorphonuclear Leukocyte and HL-60 Cell Receptors for Adhesins of Streptococcus gordonii and Actinomyces naeslundii

doi:

Figure Lengend Snippet: Bacterial overlays on nitrocellulose transfers of extracts of HL-60 cells and PMNs. Cellular extracts (20 μg of protein per lane) from HL-60 cells prior to (⊘) and during DMSO-induced differentiation as well as from PMNs were separated by SDS-PAGE and transferred to nitrocellulose. Untreated or sialidase-treated transfers were overlaid with either biotinylated S. gordonii DL1 or A. naeslundii WVU45 and washed. Bound biotinylated bacteria were detected with avidin-d-alkaline phosphatase. Lanes containing either fetuin (F), asialofetuin (AF), or thyroglobulin (TG) (5 μg per lane) were included as controls. Sizes of molecular-mass markers are indicated to the left in kilodaltons.

Article Snippet: Transfers were subsequently incubated for 1 h at room temperature with biotinylated Limax flavus agglutinin (LFA), biotinylated peanut agglutinin (PNA), or biotinylated Canavalia ensiformis agglutinin (ConA) (EY Laboratories, Inc., San Mateo, Calif.) at concentrations of 5 μg of LFA or PNA per ml or 50 μg of ConA per ml in blocking buffer.

Techniques: SDS Page, Avidin-Biotin Assay

Journal:

Article Title: Identification of Polymorphonuclear Leukocyte and HL-60 Cell Receptors for Adhesins of Streptococcus gordonii and Actinomyces naeslundii

doi:

Figure Lengend Snippet: Identification of leukosialin (CD43) and leukocyte common antigen (CD45) as PMN receptors for S. gordonii DL1 and A. naeslundii WVU45. (A) An extract of biotinylated PMNs and immunoprecipitates prepared with anti-CD43 or anti-CD45 MAbs were subjected to SDS-PAGE and transferred to nitrocellulose. Avidin-d-alkaline phosphatase was used to detect biotinylated proteins present in the extract (PMN) and immunoprecipitates (anti-CD43 or anti-CD45). (B) Immunoprecipitates (anti-CD43 or anti-CD45) from unlabeled cells were separated by SDS-PAGE and transferred to nitrocellulose. Untreated or sialidase-treated transfers were overlaid with either biotinylated S. gordonii DL1 or biotinylated A. naeslundii WVU45. Bound biotinylated bacteria were detected with avidin-d-alkaline phosphatase. Sizes of molecular-mass markers are indicated to the left in kilodaltons.

Article Snippet: Transfers were subsequently incubated for 1 h at room temperature with biotinylated Limax flavus agglutinin (LFA), biotinylated peanut agglutinin (PNA), or biotinylated Canavalia ensiformis agglutinin (ConA) (EY Laboratories, Inc., San Mateo, Calif.) at concentrations of 5 μg of LFA or PNA per ml or 50 μg of ConA per ml in blocking buffer.

Techniques: SDS Page, Avidin-Biotin Assay

Journal: PLoS Neglected Tropical Diseases

Article Title: Carbohydrate Recognition Specificity of Trans-sialidase Lectin Domain from Trypanosoma congolense

doi: 10.1371/journal.pntd.0004120

Figure Lengend Snippet: 100 μg TconTS1 and TconTS2 expressed in CHO-Lec1 cells were incubated without (-) or with (+) 4000 units EndoH f glycosidase under native (-) or denaturing (+) conditions as described under Methods. A: 10% SDS polyacrylamide gel with subsequent Coomassie Brilliant Blue staining. B: Western blot of deglycosylated TconTS, detected using anti- Strep -tag mAb. C: Concanavalin A (ConA) lectin blot using 2 μg/mL biotinylated ConA and an peroxidase conjugated avidin-biotin system (ABC-Kit, VECTASTAIN) for detection. 50 ng TconTS sample were used for ConA and Western blot analysis and 800 ng for SDS-PAGE.

Article Snippet: Concanavalin A (ConA), Sepharose and biotinylated recombinant ConA were purchased from Galab, Germany.

Techniques: Incubation, Staining, Western Blot, Strep-tag, Avidin-Biotin Assay, SDS Page