Mtb lysate

Manufactured by BEI Resources

Mtb-lysate is a biological product derived from the Mycobacterium tuberculosis (Mtb) strain. It contains a complex mixture of Mtb cellular components, including proteins, lipids, and other biomolecules, obtained through a controlled lysis process. The core function of Mtb-lysate is to provide a representative sample of Mtb constituents for research and analytical applications.

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Lab products found in correlation

Ham s f12 medium, by Thermo Fisher Scientific (1 mentions) Normocin, by InvivoGen (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Facsaria 2, by BD (1 mentions) Onestep rt pcr buffer, by Qiagen (1 mentions) Live dead fixable aqua stain, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Mtb whole cell lysate (WCL) from strain H37Rv (2 citations) Mtb H37Rv lysate (2 citations)

3 protocols using mtb lysate

Tonsil Organoid Culture and Treatment

Cited 1 time

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Tonsil organoids were established as previously described (68 (link), 69 (link)). Briefly, whole tonsils (overall healthy, without obvious signs of inflammation) were collected in saline after surgery and then immersed in an antimicrobial bath of Ham’s F12 medium (Gibco) containing Normocin (InvivoGen), penicillin, and streptomycin for 1 h at 4 °C for decontamination. Tonsils were then briefly rinsed with PBS and manually disrupted into a suspension by processing through a 100 μm strainer with a syringe plunger and cryopreserved. Frozen cells were thawed, washed, enumerated, and then plated (6 × 10⁶ cells in 100 μL per well) into permeable (0.4 μm pore size) membranes placed in standard 12-well tissue-culture plates. Organoids were cultured for 7 d at 37 °C, 5% CO₂ with or without Mtb-lysate (10 μg/mL, BEI Resources) or LAIV (1 μL per well, an equivalent of 1.6 × 10⁴ to 1.6 × 10⁵ fluorescent focus units per strain; FluMist Quadrivalent, Medimmune). Harvested cells were washed followed by flow cytometry analysis.

Guo J., Chowdhury R.R., Mallajosyula V., Xie J., Dubey M., Liu Y., Li J., Wei Y.L., Palanski B.A., Wang C., Qiu L., Ohanyan M., Kask O., Sola E., Kamalyan L., Lewis D.B., Scriba T.J., Davis M.M., Dodd D., Zeng X, & Chien Y.H. (2024). γδ T cell antigen receptor polyspecificity enables T cell responses to a broad range of immune challenges. Proceedings of the National Academy of Sciences of the United States of America, 121(4), e2315592121.

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Stimulation and Phenotypic Analysis of PBMCs

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PBMCs were thawed in complete RPMI 1640 medium at 2×10⁶ cells per ml and recovered 12 hours before stimulation. PBMCs were stimulated with an Mtb-lysate (10μg/ml) (obtained from BEI Resources) or plate-bound anti-CD3 antibody (10μg/ml) for 14 hours, and then stained with anti-CD3 (UCHT1), anti-TCRδ (5A6.E9), anti-CD8 (RPA-T8), anti-CD19 (HIB19), and anti-CD69 (FN50) antibodies. All antibodies were purchased from BioLegend. Dead cells were stained using the LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific).

Chowdhury R.R., Valainis J.R., Dubey M., von Boehmer L., Sola E., Wilhelmy J., Guo J., Kask O., Ohanyan M., Sun M., Huang H., Huang X., Nguyen P.K., Scriba T.J., Davis M.M., Bendall S.C, & Chien Y.H. (2023). NK-like CD8+ γδ T cells are expanded in persistent Mycobacterium tuberculosis infection and chronic inflammation. Science immunology, 8(81), eade3525.

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Single-cell TCR sequencing of M.tb-specific CD8+ T cells

Cited 3 times

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Single cell TCR sequencing was performed as described previously(36 (link), 37 (link)). Briefly, cryopreserved PBMCs from 12 healthy adolescents with evidence of M.tb infection (QuantiFERON+ and/or TST+) were thawed, rested for 6 hours, and stimulated for 12 hours with M.tb lysate (10μg/mL, BEI Resources) in the presence of anti-CD49d antibody (1μg/mL), and anti-CD154-PE (10μL/mL). Cells were stained with LIVE/DEAD Fixable Aqua Stain (ThermoFisher Scientific) and then with antibodies (Supplementary Table 1). Single, activated (i.e. CD69⁺CD137⁺ and/or CD69⁺CD154⁺) TCRαβ⁺ CD8⁺ cells were sorted by FACS (BD FACS Aria-II) into 96 well plates containing One-Step RT-PCR buffer (Qiagen). TCRαβ sequences were amplified using a panel of TCRαβ primers and further amplified in a nested PCR before sequencing on a MiSeq (Illumina) instrument, as described previously(36 (link), 37 (link)). TCR sequences from 6 of these 12 adolescents were published recently (38 (link)), as indicated in Supplementary Table 2.

Suliman S., Murphy M., Musvosvi M., Gela A., Meermeier E.W., Geldenhuys H., Hopley C., Toefy A., Bilek N., Veldsman A., Hanekom W.A., Johnson J.L., Boom W.H., Obermoser G., Huang H., Hatherill M., Lewinsohn D.M., Nemes E, & Scriba T.J. (2019). MR1-Independent Activation of Human Mucosal-Associated Invariant T Cells by Mycobacteria. The Journal of Immunology Author Choice, 203(11), 2917-2927.

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