Antibodies specific for the C-terminal region of human NMIIB (used at 1:1000) were generated in rabbits according to the method of (92 (link)). Recombinant GFP antibodies (used at 1:1000 dilution for Western blot and 1:200 for immunofluorescence) were prepared in rabbits as described (24 (link)). Rabbit polyclonal anti-Aurora-B antibodies (ab2254, 1:1000 for Western blot and 1:200 for immunofluorescence), mouse monoclonal anti-NMIIB (ab684, 1:200), and rat monoclonal anti-tubulin (ab6160, 1:200) were purchased from Abcam. Mouse monoclonal anti-FLAG (F3165, 1:1000) was purchased from Sigma-Aldrich. Mouse monoclonal anti-Phospho-Threonine (catalog no. 9386, clone 42H4, 1:500) was purchased from Cell Signaling Technology. Horseradish peroxidase–conjugated secondary antibodies, donkey anti-rat-IgG conjugated to Alexa Fluor 555, goat anti-mouse-IgG conjugated to Alexa Fluor 488, goat anti-rabbit-IgG conjugated to Cy5, and goat anti-mouse conjugated to Cy5 were from Jackson ImmunoResearch Laboratories.
Goat anti mouse igg conjugated to alexa fluor 488
Manufactured by Jackson ImmunoResearch
Sourced in United States
Goat anti-mouse-IgG conjugated to Alexa Fluor 488 is a secondary antibody reagent. The antibody is specific for mouse immunoglobulin G (IgG) and is labeled with the Alexa Fluor 488 fluorescent dye.
Automatically generated - may contain errors
Lab products found in correlation
Rat monoclonal anti tubulin, by Abcam (1 mentions)
Mouse monoclonal anti flag f3165, by Merck Group (1 mentions)
Ab2254, by Abcam (1 mentions)
Ab6160, by Abcam (1 mentions)
Lsm 780, by Zeiss (1 mentions)
Mouse monoclonal anti flag, by Merck Group (1 mentions)
Apotome optical sectioning, by Zeiss (1 mentions)
Vectashield, by Merck Group (1 mentions)
Axiocam mrm rev 3 camera, by Zeiss (1 mentions)
P7949, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
A9647, by Merck Group (1 mentions)
Axio observer z1 microscope, by Zeiss (1 mentions)
4 protocols using goat anti mouse igg conjugated to alexa fluor 488
1
Antibody Generation and Characterization
2
Zika Virus Neutralization and ADE Assay
3
Immunofluorescence Staining of ER and FLAG-tagged CALR
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Prior to immunofluorescence staining cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100/phosphate-buffered saline and blocked with 10% goat serum/phosphate-buffered saline. To visualize the ER chicken anti Bip IgY28 (link) was used as primary and goat anti-chicken IgG conjugated to Alexa647 (Life Technologies) as secondary antibodies. FLAG-tagged CALR variants were detected using mouse monoclonal anti-FLAG as primary (Sigma-Aldrich, St Louis, MO, USA), and goat anti mouse IgG conjugated to Alexa Fluor488 (Jackson Immuno Research Laboratories, West Grove, PA, USA) as secondary antibodies. Nuclei were counterstained with Hoechst 33342 (2μg/ml in phosphate-buffered saline). The stained samples were analysed by the laser scanning confocal microscopy system (LSM 780; Carl Zeiss, Jena, Germany) with a Plan-Apo-chromat × 60 oil immersion lens (NA 1.4).
Materials and methods on the KISMET method (cell lines, kinase inhibitors, statistics, single-dose–response values, combination of two off-target data sets, selection of kinases used for KISMET and algorithm for the calculation of the kinase rank) can be found in theSupplementary Information .
Materials and methods on the KISMET method (cell lines, kinase inhibitors, statistics, single-dose–response values, combination of two off-target data sets, selection of kinases used for KISMET and algorithm for the calculation of the kinase rank) can be found in the
4
Immunofluorescence Imaging of Oocytes
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Zona pellucida removal was performed by incubating oocytes briefly in acidic MEM-compatible medium (116.4 mM NaCl, 5.4 mM KCl, 10 mM HEPES, 1 mM NaH2PO4, 0.8 mM MgSO4, pH 1.5). Oocytes were then fixed in 4% paraformaldehyde (Sigma-Aldrich, P6148) in PBS at room temperature for 30 min, permeabilized in PBS with 0.1% Triton X-100 (Fisher Scientific, BP151–500) for 15 min at room temperature, and blocked in PBS with 0.1% BSA (Sigma-Aldrich, A9647) and 0.01% Tween-20 (Sigma-Aldrich, P7949) for 1 h at room temperature. Anti-Myc primary antibody (Thermo Fisher, MA1–21316) was used at a final concentration of 4 μg/ml. After overnight incubation with anti-Myc antibody, oocytes were washed three times in PBS with 0.1% BSA and 0.1% Tween-20 before incubation with 7.5 μg/ml goat anti-mouse IgG conjugated to Alexa Fluor 488 (Jackson ImmunoResearch) for 1 h at room temperature. Following three washes in PBS with 0.1% BSA and 0.1% Tween-20, oocytes were mounted in VectaShield containing 0.75 μg/ml 4′-6′-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, D9542). Imaging was performed using a Zeiss Axio Observer Z1 microscope with AxioCam MRm Rev3 camera and ApoTome optical sectioning (Carl Zeiss, Inc.).
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