The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
IP: Use at an assay dependent dilution (PMID 18722174).
WB: Use at a concentration of 1 - 5 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 93 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionDNA-binding protein involved in cell cycle control. May act as a transcription activator. Component of the PRP19-CDC5L complex that forms an integral part of the spliceosome and is required for activating pre-mRNA splicing.
Tissue specificityUbiquitously expressed in both fetal and adult tissues.
Involvement in diseaseA chromosomal aberration involving CDC5L is found in multicystic renal dysplasia. Translocation t(6;19)(p21;q13.1) with USF2.
Sequence similaritiesBelongs to the CEF1 family. Contains 2 HTH myb-type DNA-binding domains.
Post-translational modificationsPhosphorylated on serine and threonine residues. Phosphorylation on Thr-411 and Thr-438 is required for CDC5L-mediated mRNA splicing. Has no effect on subcellular location nor on homodimerization. Phosphorylated in vitro by CDK2. Phosphorylation enhances interaction with PPP1R8.
Cellular localizationNucleus. Nucleus speckle. Cytoplasm. May shuttle between cytoplasm and nucleus.
ICC/IF image of ab31779 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31779, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab31779 staining in human stomach, showing staining of the glanular cells (in brown). Paraffin embedded lower stomach tissue was incubated with ab31779 (1:60 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab31779 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
References for Anti-CDC5L antibody (ab31779)
This product has been referenced in:
van Maldegem F et al. CTNNBL1 facilitates the association of CWC15 with CDC5L and is required to maintain the abundance of the Prp19 spliceosomal complex. Nucleic Acids Res43:7058-69 (2015).
Read more (PubMed: 26130721) »
David CJ et al. The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65-Prp19 complex. Genes Dev25:972-83 (2011).
Read more (PubMed: 21536736) »