Mouse Anti-LAM Recombinant Antibody (clone CS-35) (CAT#: PABW-158)

Figure 1 Analysis of existing LAM mimotopes.

F-J Mimotope peptides or their scramble controls were conjugated to KLH and mimotope behavior determined by ELISA using the mouse monoclonal CS-35 anti-LAM antibody. K-M Mice (n = 5 per peptide) were vaccinated with PBS, KLH alone, or mimotope peptides or scramble controls conjugated to KLH and sera collected after 4 vaccinations.

Shin, H. J., Franco, L. H., Nair, V. R., Collins, A. C., & Shiloh, M. U. (2017). A baculovirus-conjugated mimotope vaccine targeting Mycobacterium tuberculosis lipoarabinomannan. PloS one, 12(10), e0185945.

Figure 2 LAM associates with lipid rafts and CD3 on human CD4⁺ T cells and upregulates GRAIL upon activation with anti-CD3/CD28.

(A) Human CD4+ T cells were incubated with LAM for 1 h at 37°C. After incubation with LAM, cells were incubated with Alexa Fluor 647–conjugated CT-B (1 μg/ml) for 20 min on ice, washed extensively, labeled with anti-LAM mAb (clone Cs35), followed by Alexa Fluor 488–conjugated anti-mouse IgG, and fixed with 1% paraformaldehyde (upper panels). After incubation with LAM, cells were labeled on ice with anti-LAM mAb (clone Cs35), followed by Alexa Fluor 488–conjugated anti-mouse IgG. Then, Alexa Fluor 647–conjugated anti-CD3 mAb was used to label the CD3-TCR complex (lower panels). Cells were visualized and images were merged using a Leica DM16000B confocal microscope under oil immersion objective (63×).

Sande, O. J., Karim, A. F., Li, Q., Ding, X., Harding, C. V., Rojas, R. E., & Boom, W. H. (2016). Mannose-capped lipoarabinomannan from Mycobacterium tuberculosis induces CD4+ T cell anergy via GRAIL. The Journal of Immunology, 196(2), 691-702.

Figure 3 LAM induces anergy in P25 CD4+ T cells.

(B) P25 TCR-Tg CD4⁺ T cells (1 × 10⁶), pretreated with LAM for 1 h, were stained with anti-LAM mAb or with an isotype control mAb before priming (left panel) or 5 d after priming and before restimulation (right panel) and analyzed by flow cytometry.

Sande, O. J., Karim, A. F., Li, Q., Ding, X., Harding, C. V., Rojas, R. E., & Boom, W. H. (2016). Mannose-capped lipoarabinomannan from Mycobacterium tuberculosis induces CD4+ T cell anergy via GRAIL. The Journal of Immunology, 196(2), 691-702.

Figure 1 Mouse Anti-LAM Recombinant Antibody (PABW-158) in SDS-PAGE

SDS-PAGE analysis of PABW-158 in non-reduced (Lane 1) and reduced (Lane 2) conditions.

Figure 2 Mouse Anti-LAM Recombinant Antibody (PABW-158) in HPLC

HPLC analysis of PABW-158 shows the purity is>95%.

Figure 3 Anti-LAM Recombinant Antibody (PABW-158) in WB

Western blot analysis was performed by loading 0.5 µg (lane 1), 1 µg(lane 2), and 2 µg (lane 3) Native M. Tuberculosis LAM antigen (DAGA-168) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least 1 hour. Membranes were probed with Anti-LAM Recombinant Antibody (PABW-158) at a dilution of 1:500 overnight at 4°C on a rocking platform. Then probed with Goat anti-Mouse IgG HRP secondary antibody at a dilution of 1:2,000 for one hour. Chemiluminescent was detected.
Lane 1: Reducing antigen (0.5 μg)
Lane 2: Reducing antigen (1 μg)
Lane 3: Reducing antigen (2 μg)

Figure 4 Anti-LAM Recombinant Antibody (PABW-158) in ELISA

ELISA analysis of PABW-158 was performed by coating with Native M. Tuberculosis LAM antigen (DAGA-168). Then blocked with BSA, and incubated with Anti-LAM Recombinant Antibody (PABW-158). The HRP-conj μgated goat anti-mouse IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 5 Anti-LAM Recombinant Antibody (PABW-158, lot#CB2206LX02P6) in SDS-PAGE

SDS-PAGE analysis of PABW-158 in non-reduced and reduced conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 130-180 kDa.
Lane M: Protein Marker
Lane 1: Reducing (1.5 μg)
Lane 2: Non-Reducing (1.5 μg)

Figure 6 Anti-LAM Recombinant Antibody (PABW-158, lot#CB2206LX02P6) in SEC-HPLC

HPLC analysis of PABW-158 shows the purity is>95%.