Journal: Clinical and Vaccine Immunology

Article Title: Production and Characterization of Monoclonal Antibodies against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00353-06

Figure Lengend Snippet: FIG. 1. Immunoblot analysis of mAbs 8G2 and 13E1 shows reac- tivity with the E. coli-expressed recombinant protein as well as the native M. avium subsp. paratuberculosis 35-kDa protein. Shown are three identical immunoblots that were exposed to the antibodies indi- cated beneath each filter. Note that anti-MBP (-MBP) is a mAb to the MBP affinity tag that does not detect any M. avium subsp. paratu- berculosis proteins whereas 8G2 and 13E1 detect only the E. coli- expressed MBP-MAP2121c gene product fusion protein and the MAP2121c gene product in M. avium subsp. paratuberculosis. Lanes: 1, MBP-LacZ; 2, MBP-MAP2121c gene product fusion protein; 3, M. avium subsp. paratuberculosis whole-cell lysate. Kilodalton size mark- ers are shown in the left margin and indicated by corresponding notches between the blots.

Article Snippet: Both mAbs reacted to all three M. avium subsp. paratuberculosis strains, which included K-10 and the type strain, ATCC 19698, as well as a human isolate (Fig. 2A and B, lanes 5, 10, and 12).

Techniques: Western Blot, Recombinant

Journal: Clinical and Vaccine Immunology

Article Title: Production and Characterization of Monoclonal Antibodies against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00353-06

Figure Lengend Snippet: FIG. 2. MMP mAbs are not subspecies specific for M. avium subsp. paratuberculosis. Two identical immunoblots were probed with 13E1 (A) and 8G2 (B). In all lanes, 0.6 g of a whole-cell lysate was loaded. Lanes: 1, protein standards; 2, M. avium subsp. silvaticum; 3, M. scrofu- laceum; 4, M. abscessus; 5, M. avium subsp. paratuberculosis K-10; 6, M. avium subsp. avium (TMC702); 7, M. bovis (strain 95-1315); 8, M. phlei; 9, M. bovis BCG; 10, M. avium subsp. paratuberculosis ATCC 19698; 11, M. avium subsp. avium (TMC715); 12, M. avium subsp. paratuber- culosis (isolate Linda); 13, M. intracellulare; 14, M. kansasii. Sizes in kilodaltons of protein standards are indicated in the left margin.

Article Snippet: Both mAbs reacted to all three M. avium subsp. paratuberculosis strains, which included K-10 and the type strain, ATCC 19698, as well as a human isolate (Fig. 2A and B, lanes 5, 10, and 12).

Techniques: Western Blot

Journal: Clinical and Vaccine Immunology

Article Title: Production and Characterization of Monoclonal Antibodies against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00353-06

Figure Lengend Snippet: FIG. 4. MMP was present in the membrane-enriched fractions of five M. avium subsp. paratuberculosis isolates. The resulting immuno- blot was exposed to mAb 8G2. This antibody labeled a 35-kDa protein in the membrane-enriched fractions of the five M. avium subsp. para- tuberculosis isolates. Sizes in kilodaltons of protein standards are indi- cated in the left margin.

Article Snippet: Both mAbs reacted to all three M. avium subsp. paratuberculosis strains, which included K-10 and the type strain, ATCC 19698, as well as a human isolate (Fig. 2A and B, lanes 5, 10, and 12).

Techniques: Membrane, Labeling

Journal: Clinical and Vaccine Immunology

Article Title: Production and Characterization of Monoclonal Antibodies against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00353-06

Figure Lengend Snippet: FIG. 5. Immunofluorescent-antibody staining of 48-h M. avium subsp. paratuberculosis-infected BoMac macrophages with Alexa Fluor 488- labeled 8G2 and 13E1. Each four-panel image shows cells visualized by using DIC, DAPI, Alexa Fluor 488, and DAPI merged with Alexa Fluor 488. Both 8G2 and 13E1 Alexa Fluor 488-labeled antibodies stained intracellular M. avium subsp. paratuberculosis. Arrows in the DIC and Alexa Fluor 488 panels indicate the locations of the mycobacteria. Note that the DAPI nuclear staining is in close proximity to the Alexa Fluor 488-labeled mycobacteria. The bar indicates 5 m for all panels.

Article Snippet: Both mAbs reacted to all three M. avium subsp. paratuberculosis strains, which included K-10 and the type strain, ATCC 19698, as well as a human isolate (Fig. 2A and B, lanes 5, 10, and 12).

Techniques: Staining, Infection, Labeling