Immunogen: KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO9 protein sequence UniProt: Q84YI4, TAIR:At5g21150.
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 µg
Reconstitution: For reconstitution add 50 µl of sterile water
Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications: Immunolocalization, whole-mount (IL), Immunoprecipitation (IP), Western blot (WB)
Recommended dilutions: 5 µg of antibody per 1 gram of a fresh tissue (IP), 1 : 10 000 (WB)
Hou et al. (2021) High-throughput single-cell transcriptomics reveals the female germline differentiation trajectory in Arabidopsis thaliana. Commun Biol. 2021 Oct 1;4(1):1149. doi: 10.1038/s42003-021-02676-z. PMID: 34599277; PMCID: PMC8486858. (immunolocalization)Oliver & Martinez. (2021) Accumulation dynamics of ARGONAUTE proteins during meiosis in Arabidopsis. Plant Reprod. 2021 Nov 23. doi: 10.1007/s00497-021-00434-z. Epub ahead of print. PMID: 34812935.Sprunck et al. (2019). Elucidating small RNA pathways in Arabidopsis thaliana egg cells. http://dx.doi.org/10.1101/525956Su et al. (2017). The THO Complex Non-Cell-Autonomously Represses Female Germline Specification through the TAS3-ARF3 Module. Curr Biol. 2017 Jun 5;27(11):1597-1609.e2. doi: 10.1016/j.cub.2017.05.021.Havecker et al. (2010) The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. Plant Cell 22(2): 321-34.
AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control. Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure.A recommended whole-mount immunolocalization protocol can be found here.