Mili Antibody [K20L16] | Primary Antibodies

Application: Reactivity: Mouse

Lane 1: Mouse testis

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:100 IF1:50

Application
WB, IP, IHC, IF

Reactivity
Mouse

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
110 kDa

Datasheet & SDS

Biological Description

Specificity
Mili Antibody [K20L16] detects endogenous levels of total Mili protein.

Clone
K20L16

Synonym(s)
Piwi-like protein 2; Piwil2

Background
Mili functions as a key member of the PIWI subfamily within the Argonaute protein family, alongside Miwi and Miwi2, where it specifically associates with Piwi-interacting RNAs (piRNAs) to safeguard genomic integrity during mammalian spermatogenesis. Mili contains a conserved PAZ domain that anchors the 3' end of piRNAs and a MID domain that clamps the 5' end, coupled to a slicer-competent PIWI domain that enables endonucleolytic cleavage of complementary transcripts while facilitating interactions with Tudor-domain proteins through symmetric dimethylarginine modifications on its N-terminal tail. Primary piRNA biogenesis initiates with Mili binding long single-stranded precursors in the cytoplasm of prospermatogonia, where it licenses phased slicing that generates 5' monophosphorylated intermediates for loading onto secondary PIWI proteins, establishing a ping-pong amplification cycle that populates nuage and chromatoid bodies with transposon-targeting piRNAs. Interaction with MVH helicase unwinds precursor duplexes to expose targets, while Tudor domain-containing protein 1 (TDRD1) scaffolds Mili within cytoplasmic granules to concentrate piRNA-directed silencing machinery, coordinating mRNA deadenylation and decay of retrotransposon transcripts like LINE1 and IAP that threaten meiotic progression. Mili further engages translational initiation factors eIF4G and eIF3A to selectively repress or stabilize germ cell-specific mRNAs, integrating piRNA-guided cleavage with post-transcriptional control during germ granule maturation. Mili sustains germline stem cell self-renewal and spermatogonial differentiation by restricting transposon mobility, with peak expression from embryonic prospermatogonia through early pachytene spermatocytes reflecting stage-specific piRNA repertoire expansion. Dysregulation through Mili depletion arrests spermatogenesis at the spermatocyte stage, unleashing transposon derepression that compromises meiotic chromosome pairing and DNA methylation at retroelement loci.

Background

Mili functions as a key member of the PIWI subfamily within the Argonaute protein family, alongside Miwi and Miwi2, where it specifically associates with Piwi-interacting RNAs (piRNAs) to safeguard genomic integrity during mammalian spermatogenesis. Mili contains a conserved PAZ domain that anchors the 3' end of piRNAs and a MID domain that clamps the 5' end, coupled to a slicer-competent PIWI domain that enables endonucleolytic cleavage of complementary transcripts while facilitating interactions with Tudor-domain proteins through symmetric dimethylarginine modifications on its N-terminal tail. Primary piRNA biogenesis initiates with Mili binding long single-stranded precursors in the cytoplasm of prospermatogonia, where it licenses phased slicing that generates 5' monophosphorylated intermediates for loading onto secondary PIWI proteins, establishing a ping-pong amplification cycle that populates nuage and chromatoid bodies with transposon-targeting piRNAs. Interaction with MVH helicase unwinds precursor duplexes to expose targets, while Tudor domain-containing protein 1 (TDRD1) scaffolds Mili within cytoplasmic granules to concentrate piRNA-directed silencing machinery, coordinating mRNA deadenylation and decay of retrotransposon transcripts like LINE1 and IAP that threaten meiotic progression. Mili further engages translational initiation factors eIF4G and eIF3A to selectively repress or stabilize germ cell-specific mRNAs, integrating piRNA-guided cleavage with post-transcriptional control during germ granule maturation. Mili sustains germline stem cell self-renewal and spermatogonial differentiation by restricting transposon mobility, with peak expression from embryonic prospermatogonia through early pachytene spermatocytes reflecting stage-specific piRNA repertoire expansion. Dysregulation through Mili depletion arrests spermatogenesis at the spermatocyte stage, unleashing transposon derepression that compromises meiotic chromosome pairing and DNA methylation at retroelement loci.

References
https://pubmed.ncbi.nlm.nih.gov/19345100/ https://pubmed.ncbi.nlm.nih.gov/14736746/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%