Las af software version 1.6.0 build 1016

Manufactured by Leica

Sourced in Germany

LAS AF software version 1.6.0 build 1016 is a microscope imaging and analysis software developed by Leica. It provides a platform for acquiring, processing, and analyzing microscopic images.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594, by Thermo Fisher Scientific (2 mentions) Ab23905, by Abcam (1 mentions) T6074, by Abcam (1 mentions) Tcs sp5, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)

Close spelling variants of this product

LAS AF version 1.6.3 software LAS AF software LAS AF 1.8.2 software LAS AF LAS software

2 protocols using las af software version 1.6.0 build 1016

Immunostaining of Cell Lines

Cited 1 time

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A549, H1650, H460, Calu-6, PC9 or AALE cells were plated onto poly-D-lysine-coated eight-well glass chamber slides (5,000 cells per well) for immunostaining. The cells were fixed with 10% buffered formalin and indirect immunofluorescence was performed as described56 (link). Primary antibodies used were phospho-TBK1 (Cell signaling #5483S) or CEP170 mouse monoclonal-72-413-1 (Invitrogen #41-3200) at 1:100 dilution or 1:500 dilution of rabbit monoclonal NuMA (EP3976 Abcam #109262), Alpha tubulin (1:2,000, Sigma #T6074), Dynein intermediate chain (Abcam#ab23905) and Kif2b (Abcam #ab98214 or Novus #NBP86002), γ Tubulin (Santa Cruz #sc51715) were used at 1:100 dilution. Anti-rabbit Alexa Fluor-488 or anti- mouse Alexa Fluor-594 (Molecular Probes) was used as secondary antibody. DAPI (Vector Labs) was used to stain the nuclei. Cells were visualized with a DM16000 inverted Leica TCS SP5 tandem scanning confocal microscope with a × 63/1.40NA oil immersion objective. Images and Z-stacks were produced with three cooled photomultiplier detectors and analysed with the LAS AF software version 1.6.0 build 1016 (Leica Microsystems, Germany).

Pillai S., Nguyen J., Johnson J., Haura E., Coppola D, & Chellappan S. (2015). Tank binding kinase 1 is a centrosome-associated kinase necessary for microtubule dynamics and mitosis. Nature Communications, 6, 10072.

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Immunofluorescence Microscopy of Cell Lines

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A549, H460, MDA-MB-231 and Hs578T cells were seeded on eight-well glass chamber slides at a density of 5,000 cells per well. After indicated treatments cells were fixed with 10% buffered formalin for 20 mins or with chilled methanol at −20°C for 5 minutes, followed by cell permeabilization with PBS containing 0.25% Triton X-100 in case of formalin fixation. Cells were then blocked using 5% goat serum for an hour and incubated with primary antibody overnight. Primary antibodies used were phospho-TBK1 (1:200), Alpha tubulin (1:2,000), Gamma tubulin (1:250) and Cdc20 (1:200). Anti-rabbit Alexa Fluor-488 or anti-mouse Alexa Fluor-594 (Molecular Probes) was used as secondary antibody. DAPI was used to stain the nuclei. Cells were visualized with a DM16000 inverted Leica TCS SP8 tandem scanning confocal microscope with 63x/1.40NA oil immersion objective. Images and Z-stacks were produced with three cooled photomultiplier detectors and analyzed with the LAS AF software version 1.6.0 build 1016 (Leica Microsystems, Germany).

Maan M., Agrawal N.J., Padmanabhan J., Leitzinger C.C., Rivera-Rivera Y., Saavedra H.I, & Chellappan S.P. (2020). Tank Binding Kinase 1 modulates spindle assembly checkpoint components to regulate mitosis in breast and lung cancer cells. Biochimica et biophysica acta. Molecular cell research, 1868(3), 118929.

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