Cells were transfected in 8-well glass chambers (Millipore) and fixed with 4% paraformaldehyde 24 hours later. Cells were permeabilized with 0.1% Triton-X100 and blocked with 10% donkey serum. GFP and mCherry fluoresce was detected directly. The primary antibodies used were: SC-35 (1:300; Abcam, ab11826), PML (1:50; Santa Cruz, sc-966), coilin (1:500; Santa Cruz Biotechnology, sc-32860), B23 (1:200, Santa Cruz Biotechnology, sc-56622) Myc-tag (1:500 Cell Signaling Technologies, 71D10), HA-tag (1:250; Clone 3F10, Roche, 11867423001), FK2 (1:50; Enzo Life Sciences, BML-PW8810), V5-tag (1:300, Novus Biological, NB600–379). The secondary antibodies used were Alexa 555, 647 (1:5,000; Thermo-Fisher), and CF405S (1:1000; Biotium). Samples were mounted on ProLong Gold antifade with or without DAPI and cured before imaging on a Zeiss LSM 780 NLO microscope. Images were prepared with the Fiji software.
Sc 35
Manufactured by Abcam
Sourced in United Kingdom, United States
SC-35 is a monoclonal antibody that recognizes the SC-35 protein, a member of the serine/arginine-rich family of pre-mRNA splicing factors. This antibody is commonly used in research applications to detect and analyze the localization and expression of the SC-35 protein.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 555, by Thermo Fisher Scientific (1 mentions)
Coilin, by Santa Cruz Biotechnology (1 mentions)
Cf405s, by Biotium (1 mentions)
Ha tag, by Roche (1 mentions)
Myc tag, by Cell Signaling Technology (1 mentions)
Lsm 780 nlo microscope, by Zeiss (1 mentions)
Lsm 700, by Zeiss (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
Bcl xl, by Abcam (1 mentions)
Hrp conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Srpk2, by Merck Group (1 mentions)
Sc 35, by Merck Group (1 mentions)
4 protocols using sc 35
1
Multicolor immunofluorescence protocol
2
Immunofluorescence Staining of Bcl-xL and SC35
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded and grown on Ψ20 mm glass-bottom cell culture dishes and fixed with 0.4% paraformaldehyde. Subsequently, cells were permeabilized in 0.5% Triton X-100, blocked in 10% BSA, and incubated overnight at 4 °C with primary antibodies. The following primary antibodies were tested: Bcl-xL (1:500, Abcam), SC35(1:200, Abcam). The cells were then stained with secondary antibodies and 4′,6-diamidino-2-phenylindole (Vector Laboratories, Burlingame, CA). Fluorescence images were visualized and captured using a confocal scanning microscope (LSM700, Zeiss, Germany).
3
Quantitative Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed on total proteins from patient tissues, cultured cells and xenograft tumors tissues. The protein was separated by dodecyl sulfate, sodium salt-polyacrylamide gel electrophoresis (SDS-PAGE), then the protein expressions were detected using standard techniques for immunoassays. Antibodies to CyclinE (SAB, #29030, 1:1000 dilution), SRPK2 (Sigma-Aldrich, #HPA015522, 1:1000 dilution), p45SKP2 (Invitrogen, #323300, 1:2000 dilution), SC35 (Abcam, Cambridge, UK, #ab204916, 1:2000 dilution), E2F1 (Abcam, #ab179445, 1:2000 dilution). HRP-conjugated secondary antibodies (Invitrogen, G-21040) were used for the blotting. Controls were referred to β-actin (Cell Signaling Technology, Danvers, MA, USA; #4970, 1:5000 dilution). The expression of each protein was analyzed using biometric digital image software and recorded as integral density (ID). The final result of the ratio of target proteins were calculated as ID (per protein) / ID (β-actin) protein expression.
4
Subcellular localization of NONO and SC-35
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded on coverslips and then fixed with 4% paraformaldehyde. Immunofluorescence staining was performed to determine the subcellular localization of NONO and SC-35 with the following antibodies: NONO antibody (Abcam) and SC-35 (Sigma-Aldrich; St. Louis, MO, USA). FAM or CY3 modified FISH probes were used to detect the GPX1 pre-mRNA and mature mRNA following the manufacturer's protocol (GenePharma).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!