The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Blocking - Blocking peptide for Anti-PIWIL4 antibody (ab21869)
- First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions. - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer. - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent. - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised. - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Information available upon request.
Miwi 2 protein
Piwi like 2
Piwi like 4
Piwi like 4 (Drosophila)
PIWI like protein
Piwi like protein 4
Piwi like RNA mediated gene silencing 4
Piwi-like protein 4
FunctionPlays a central role during spermatogenesis by repressing transposable elements and prevent their mobilization, which is essential for the germline integrity. Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and govern the methylation and subsequent repression of transposons. Directly binds piRNAs, a class of 24 to 30 nucleotide RNAs that are generated by a Dicer-independent mechanism and are primarily derived from transposons and other repeated sequence elements. Associates with secondary piRNAs antisense and PIWIL2/MILI is required for such association. The piRNA process acts upstream of known mediators of DNA methylation. Participates to a piRNA amplification loop. Besides their function in transposable elements repression, piRNAs are probably involved in other processes during meiosis such as translation regulation (By similarity). May be involved in the chromatin-modifying pathway by inducing 'Lys-9' methylation of histone H3 at some loci.
Tissue specificityExpressed in testis. According to PubMed:17544373, it is ubiquitously expressed.
Sequence similaritiesBelongs to the argonaute family. Piwi subfamily. Contains 1 PAZ domain. Contains 1 Piwi domain.
Post-translational modificationsArginine methylation by PRMT5 is required for the interaction with Tudor domain-containing protein (TDRD1, TDRKH/TDRD2 and TDRD9) and subsequent localization to the meiotic nuage, also named P granule.
Cellular localizationNucleus. Cytoplasm. Probable component of the meiotic nuage, also named P granule, a germ-cell-specific organelle required to repress transposon during meiosis. PIWIL2/MILI is required for nuclear localization.
Lane 1 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027) Lane 2 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027) with Mouse PIWIL4 peptide (ab23535) at 1 µg/ml Lane 3 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027) Lane 4 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027) with Mouse PIWIL4 peptide (ab23535) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 98 kDa Observed band size : 98 kDa