Anti coilin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-Coilin is a monoclonal antibody that recognizes the coilin protein, which is a marker for Cajal bodies in the nucleus. Coilin is a protein involved in the assembly and organization of these nuclear bodies, which play a role in the processing and modification of small nuclear ribonucleoproteins (snRNPs). This antibody can be used to detect and study the localization of coilin and Cajal bodies in various cell types and tissues.

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4 protocols using anti coilin

Immunofluorescence Analysis of SUMO-2/3

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HeLa cells were grown in DMEM supplemented with 10% fetal calf serum (FCS) at 37°C with 5% CO2. LMB was added at a 10 nM final concentration from a 10 µm stock solution in 70% methanol (Sigma-Aldrich). An equivalent amount of methanol was added to control cells. The following rabbit polyclonal antibodies were used in this study: anti-SUMO-2/3 (Abcam 3742), anti-SUMO1 (Santa Cruz sc FL-101), anti-CRM1 (Santa Cruz sc-5595), anti-Histone H3 (Abcam1791), anti-p62/SQSTM1 (Santa Cruz sc-25575), anti-Ubiquitin (DAKO Z 0458), anti-Coilin (Santa Cruz sc-32860), rabbit anti-PML.⁶⁴ (link) Monoclonal mouse anti-CRM1 (BD 611 832), anti-PML (Santa Cruz sc PG-M3) and anti-p62/SQSTM1 (BD transduction Laboratories 610832) were used single or in conjunction with rabbit antibodies for double-labeling experiments.

Souquere S., Weil D, & Pierron G. (2015). Comparative ultrastructure of CRM1-Nucleolar bodies (CNoBs), Intranucleolar bodies (INBs) and hybrid PML/p62 bodies uncovers new facets of nuclear body dynamic and diversity. Nucleus, 6(4), 326-338.

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Co-IP, Western Blot, and Immunofluorescence Protocol

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Co-immunoprecipitation (Co-IP), western blotting and IF were carried out as previously described (47 (link)). The antibodies used for IP and western are: mouse monoclonal anti-FLAG (Abmart, M20008), rabbit polyclonal anti-GST (CST, 2622S), rabbit polyclonal anti-GAPDH (Abmart, P30008), mouse monoclonal anti-Coilin (Abcam, ab87913), rabbit polyclonal anti-TOE1 (BETHYL, A303-643A), rabbit polyclonal anti-DKC1 (Santa Cruz, sc-48794), anti-FLAG M2 Affinity Gel (Sigma, A220).
Primary antibodies for IF are: mouse monoclonal anti-Coilin (Abcam, ab87913), rabbit polyclonal anti-TOE1 (BETHYL, A303-643A), rabbit polyclonal anti-DKC1 (Santa Cruz, sc-48794), mouse polyclonal anti-DKC1 (Santa Cruz, sc-373956), rabbit polyclonal anti-Coilin (Santa Cruz, H-300). Secondary antibodies include DyLight488 goat anti-rabbit IgG (Liankebio, LK-GAR4882), Dylight488 goat anti-mouse IgG (Liankebio, LK-GAR4881), Alexa Fluor^® 555 Donkey Anti-Mouse IgG (Thermo Fisher Scientific, A-31570), Alexa Fluor^® 555 Donkey Anti-Rabbit IgG (Thermo Fisher Scientific, A-31572).

Deng T., Huang Y., Weng K., Lin S., Li Y., Shi G., Chen Y., Huang J., Liu D., Ma W, & Songyang Z. (2018). TOE1 acts as a 3′ exonuclease for telomerase RNA and regulates telomere maintenance. Nucleic Acids Research, 47(1), 391-405.

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Quantitative Protein Expression Analysis

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The treated cells were lysed following a standard protocol. The protein expression was assessed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting analysis. For ALK signaling experiments, confluent cell monolayer of the H3122 cells were serum-free starved 24 h and then exposed to 100 μg/ml heparin for 30 min. The anti-NHERF1 antibody was purchased from BD Transduction Laboratories (BD Biosciences, San Diego, CA, USA). The anti-GAPDH, anti-ERK, anti-p-ERK and anti-Coilin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-AKT, anti-p-AKT (Ser473), anti-ALK, anti-p-ALK (Tyr1604) antibodies were from Cell Signaling Technology (Danvers, MA, USA). The β-action antibody was obtained from ABclonal Biotechnology (Wuhan, Hubei, China). The protein expression levels were semi-quantified by densitometry of the western blot bands using the ImageJ2 software.

Yang F., Hu M., Chang S., Huang J., Si Y., Wang J., Cheng S, & Jiang W.G. (2020). Alteration in the sensitivity to crizotinib by Na+/H+ exchanger regulatory factor 1 is dependent to its subcellular localization in ALK-positive lung cancers. BMC Cancer, 20, 202.

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RNA Pulldown Assay Protocol

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RNA pulldowns were conducted as previously described (Poole et al., 2016 (link)). The following antibodies were used: anti-coilin (Santa Cruz Biotechnology), anti-fibrillarin (Santa Cruz Biotechnology), anti-SMN (Abcam), and anti-β-tubulin (Santa Cruz Biotechnology). Membranes were visualized and quantified using a ChemiDoc (Bio-Rad) with QuantityOne software. Data were imported to Microsoft Excel, and statistical significance determined using the Student's t-test.

Poole A.R., Vicino I., Adachi H., Yu Y.T, & Hebert M.D. (2017). Regulatory RNPs: a novel class of ribonucleoproteins that potentially contribute to ribosome heterogeneity. Biology Open, 6(9), 1342-1354.

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