Cells were fixed with 3.7% formaldehyde 20 mM HEPES in PBS for 10min. This step was skipped when working with Jurkat cells as they were fixed during the setup of the cells. The formaldehyde was quenched with 10 mM Tris in PBS for 10mins, and the cells were then permeabilised for 5mins with PBS containing 0.05% Tx-100. The cells were then blocked with 3% BSA in PBS for 20mins and then stained with primary antibodies (Ticilimumab, anti-LRBA (Atlas Antibodies), anti-Rab5, anti-Rab7, anti-Rab9 (all Cell Signaling Technology), anti-Rab11 (Invitrogen)) in 3mg/ml BSA in PBS for 1hr at RT. Next, goat anti-human IgG-AlexaFluor 546, donkey anti-rabbit IgG-AlexaFluor 488 (both Invitrogen), Hoechst 33342 (Thermo Fisher Scientific) and 2.5 μM CellTrace Violet (Molecular Probes) in 3mg/ml BSA in PBS were added to the cells for 45mins. All steps up to and including the 3% BSA block were carried out on ice, all the following steps were at RT, and the cells were washed 3 times with PBS between each step.
Ticilimumab
Manufactured by Atlas Antibodies
Ticilimumab is a monoclonal antibody produced by Atlas Antibodies. It is a laboratory reagent used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti rab5, by Cell Signaling Technology (2 mentions)
Anti rab9, by Cell Signaling Technology (2 mentions)
Goat anti human igg alexafluor 546, by Thermo Fisher Scientific (2 mentions)
Celltrace violet, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Anti rab11, by Thermo Fisher Scientific (2 mentions)
Anti rab7, by Cell Signaling Technology (2 mentions)
Anti lrba, by Atlas Antibodies (2 mentions)
Anti lc3a b, by Cell Signaling Technology (1 mentions)
2 protocols using ticilimumab
1
Immunofluorescence staining of fixed cells
2
Multicolor Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were fixed with 3.7% formaldehyde 20 mM HEPES in PBS for 10 min. This step was skipped when working with Jurkat cells as they were fixed during the set‐up of the cells. The formaldehyde was quenched with 10 mM Tris in PBS for 10 min, and the cells were then permeabilized for 5 min with PBS containing 0.05% Triton‐X‐100. The cells were then blocked with 3% BSA in PBS for 20 min and then stained with primary antibodies (ticilimumab and anti‐LRBA (Atlas Antibodies); anti‐Rab5, anti‐Rab7 and anti‐Rab9 (all from Cell Signaling Technology); anti‐Rab11 (Invitrogen or Atlas Antibodies) and anti‐LC3A/B (Cell Signaling Technology)) in 3 mg/ml BSA in PBS for 1 h at RT. Next, goat anti‐human IgG‐Alexa Fluor 546, donkey anti‐rabbit IgG‐Alexa Fluor 488 (both Invitrogen), Hoechst 33342 (Thermo Fisher Scientific) and 2.5 µM CellTrace Violet (Molecular Probes) in 3 mg/ml BSA in PBS were added to the cells for 45 min. All steps up to and including the 3% BSA block were carried out on ice, all the following steps were at RT, and the cells were washed 3 times with PBS between each step.
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