We used the following primary antibodies: rabbit polyclonal antibodies against SANS (RpAb) (29 (link),37 (link)), anti-SANS (mouse, monoclonal antibody; MmAb) (sc-514418, Santa Cruz), anti-FLAG (F1804, Sigma Aldrich), anti-PRPF6 (sc-48786, Santa Cruz), anti-HA (Roche), anti-GFP (ab6556, Abcam), anti-PRPF31 (PAB7154, Abnova), anti-hSNU114 (38 (link)), anti-Coilin (sc-55594, Santa Cruz), anti-SC35 (sc-53518, Santa Cruz), anti-SON (ATLAS, HPA023535), anti-SF3B1 (MmAb) (sc-514655, Santa Cruz), anti-SF3B1 (RmAb) (Will et al. 2001), anti-U5 52K (39 (link)), anti-SART1 (PA556663, Thermo Fisher) and anti-PRPF38 (40 (link)). Secondary antibodies were conjugated to the following fluorophores: Alexa488 (donkey-anti-rabbit, A21206, Molecular Probes,), CF 488 (donkey-anti-guinea pig, 20169-1, Biotrend,), Alexa488 (donkey-anti-mouse, A21202, Molecular Probes), Alexa 555 (donkey-anti-mouse, A31570, Molecular Probes), Alexa 568 (donkey-anti-rabbit, A10043, Invitrogen,) CF640 (donkey-anti-goat, 20179-1, Biotrend), Alexa 680 (donkey-anti-goat, A21084, Molecular Probes), Alexa 680 (goat-anti-rat, A21096, Molecular Probes) Alexa 680 (donkey-anti-rabbit, A10043), IRDye 800 (donkey-anti-mouse, 610–732-124, Rockland), Abberior STAR Orange (goat-anti-mouse, Abberior,), and Abberior STAR Red (goat-anti-Rabbit). Nuclear DNA was stained with DAPI (4’,6-diamidino-2-phenylindole) (1 mg/ml) (Sigma-Aldrich).
Protocol detail
Comprehensive Immunolabeling Protocol for RNA Splicing
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anti-FLAG
F1804
anti-PRPF6
sc-48786
anti-HA
ab6556
PAB7154
sc-53518
sc-514655
A21206
Alexa488
A21202
Alexa 555
A31570
Alexa 568
Alexa 680
IRDye 800
Alexa 568
Antibodies
Dapi
Donkey
Goat
Guinea pig
Irdye 800
Molecular probes
Monoclonal antibody
Mouse
Rabbit
Corresponding Organization :
Other organizations : Johannes Gutenberg University Mainz, Max Planck Institute for Biophysical Chemistry, Mashhad University of Medical Sciences, Reference Institute for Bioanalytics
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Variable analysis
independent variables
- Primary antibodies: rabbit polyclonal antibodies against SANS (RpAb), anti-SANS (mouse, monoclonal antibody; MmAb), anti-FLAG, anti-PRPF6, anti-HA, anti-GFP, anti-PRPF31, anti-hSNU114, anti-Coilin, anti-SC35, anti-SON, anti-SF3B1 (MmAb), anti-SF3B1 (RmAb), anti-U5 52K, anti-SART1, and anti-PRPF38
- Secondary antibodies conjugated to fluorophores: Alexa488, CF 488, Alexa488, Alexa 555, Alexa 568, CF640, Alexa 680, Alexa 680, Alexa 680, IRDye 800, Abberior STAR Orange, and Abberior STAR Red
- Not explicitly mentioned
- Nuclear DNA stained with DAPI
- Positive controls: Not specified
- Negative controls: Not specified
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