Anti Mycobacterium tuberculosis, L-alanine dehydrogenase (Ald)
Mouse monoclonal antibody
Cat.No. HBT 10 (WHO no. IT-46)
Preparation: Protein-A/G purified
Content: Available in 200 µL and 1 mL
Concentration: 1 mg/mL
Solvent: 0.01 M phosphate buffer, pH 7.4, with 0.5 M NaCl and 15 mM sodium azide
Storage: In the dark at 4-8ºC
L-alanine dehydrogenase (Ald) antigen from Mycobacterium tuberculosis.
Culture filtrat from Mycobacterium tuberculosis H37Rv
HBT 10 is specific for a 40 kDa band from short term culture of Mycobacterium tuberculosis in Western blotting after SDS-PAGE. The 40 kDa band was identified as the protein L-alanine dehydrogenase (Ald). Interspecies analyses with HBT 10 showed reacts with M. tuberculosis and M. marinum but not M. bovis BCG (3).
HBT 10 recognizes the 12 aa SAITDADFKAAG (aa 45-56) region of Ald of Mycobacterium tuberculosis (4).
HBT 10 reacts specifically with Ald of Mycobacterium tuberculosis. Hutter et al (4) has demonstrated that HBT 10 does not react with pyridine nucleotide transhydrogenase (PNT) of Mycobacterium tuberculosis, which has previously been suggested by Delforge et al. (5).
HBT 10 can be used in immunoblotting, after 1- and 2-dimensional gel electrophoresis (3,4,6,7,8).
| Method | Usability | Dilution guideline | References |
| ELISA | Yes | ||
| Immunoblotting | Yes | 3,4,6,7,8 | |
| Immunohistochemistry | Not determined |
1. Worsaae A, Ljungqvist L, Heron I (1988) Monoclonal antibodies produced in BALB.B10 mice define new antigenic determinants in culture filtrate preparations of Mycobacterium tuberculosis. J Clin Microbiol 26:2608-2614.
2. Andersen AB, Andersen P, Ljungqvist L (1992) Structure and function of a 40,000-molecular-weight protein antigen of Mycobacterium tuberculosis. Infect Immun 60:2317-2323.
3. Ljungqvist L, Worsaae A, Heron I (1988) Antibody response against Mycobacterium tuberculosis in 11 strains of inbred mice: Novel monoclonal antibody specificities generated by fusion, using spleens from BALB.B10 and CBA/J mice. Infect Immun 56:1994-1998.
4. Hutter B, Singh M (1998) Host vector system for high-level expression and purification of recombinant, enzymatically active alanine dehydrogenase of Mycobacterium tuberculosis. Gene 212:21-29.
5. Delforge D, Depiereux E, Bolle XD, Feytmans E, Remacle J (1993) Similarities between alanine dehydrogenase and the N-terminal part of pyridine nucleotide transhydrogenase and their possible implication in the virulence mechanism of Mycobacterium tuberculosis. Biochem Biophys Res Commun 190:1073-1079.
6. Bardou F, Quémard A, Dupont M, Horn C, Marchal G, Daffé M (1996) Effects of isoniaazid on ultrastructure of Mycobacterium aurum and Mycobacterium tuberculosis and on production of secreted proteins. Antimicrob Agents Chemother 40:2459-2467.
7. Weldingh K, Rosenkrands I, Jacobsen S, Rasmussen PB, Elhay MJ, Andersen P (1998) Two-dimensional electrophoresis for Analysis of Mycobacterium tuberculosis culture filtrate and purification and characterization of six novel proteins. Infect Immun 66:3492-3500.
8. Rosenkrands I, Weldingh K, Jacobsen S, Hansen CV, Florio W, Gianetri I, Andersen P (2000) Mapping and identification of Mycobacterium tuberculosis proteins by two-dimensional gel electrophoresis, microsequencing and immunodetection. Electrophoresis 21:935-948.
