Anti-SE | Serrate RNA effector molecule (rabbit antibody)

Product no: AS09 532A

AS09 532A  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana, Malus domestica,, Nicotiana benthamina, Nicotiana tabacum

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  • Product Info
  • Immunogen:

    KLH-conjugated synthetic peptide chosen from Arabidopsis thaliana serrate protein sequence Q9ZVD0, At2g27100

    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: Chromatin Immunoprecipitation (ChIP), Immunolocalization (IL), RNA Immunoprecipitation (RIP), Western blot (WB)
    Recommended dilution: 1 : 500 (IL), 1 : 1000 (WB)
    Expected | apparent MW:

    81 | 80 kDa

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Malus domestica, Nicotiana benthamina, Nicotiana tabacum
    Predicted reactivity: Saccharum hybrid cultivar NCo 376, Zea mays

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example

    western blot using anti-serrate antibodies

    25-30 µg of total protein from Arabidopsis thaliana rosette leaves was extracted with extraction buffer containing: 100 mM Tris HCl pH 7.5, 10 % sucrose, 5 mM EDTA, 5 mM EGTA, 300 mM NaCl, 0.75 % Triton X100, 0.15 % SDS, 1 mM DTT, 1x Complete Mini EDTA-free protease inhibitor (Roche) or 7.5 % nuclear fraction obtained according to the protocol from Raczyńska et al. 2014, were separated on 10 % SDS/PAGE using semi-dry transfer and blotted 1 h to PVDF. Blots were blocked with 2.5 % milk in PBS/T for overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1: 250 in PBS-T for 1 h at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit HRP conjugated, AS09 602, Agrisera) in 1: 5000 dlution for 1 h at RT with agitation. The blot was washed as above and developed for 5 minutes with ECL according to manufacturer's instructions. Exposure time was 600 seconds.

    Courtesy of M.Sc. Mateusz Bajczyk, Department of Gene Expression, Adam Mickiewicz University, Poland
    Application examples:

    Reactant: Arabidopsis thaliana (Thale cress)

    Application: Proximity Ligation Assay

    Pudmed ID: 36087009

    Journal: Plant Cell

    Figure Number: 1A,B

    Published Date: 2022-11-29

    First Author: Stepien, A.

    Impact Factor: 7.018

    Open Publication

    AtPRP40 regulates the association of SE with RNAPII. A, Close proximity of AtPRP40b and SE in the cell nucleus (first image, magenta signals) analyzed by PLA in view of SE nuclear localization (second image, green signals). DNA was stained with Hoechst (blue). Scale bar = 2.5 µm. B, Close proximity of SE and RNAPII phosphorylated at CTD Ser5 or Ser2 in WT, prp40ab, and se-2 plants (first columns, magenta signals) in view of P-Ser5-RNAPII or P-Ser2-RNAPII nuclear localization (second columns, green signals) detected by PLA. DNA was stained with Hoechst (blue). Scale bar = 2.5 µm. C, Close proximity of AtPRP40b and RNAPII phosphorylated at CTD Ser5 or Ser2 in WT, se-2, and prp40ab plants (first columns, magenta signals) in view of P-Ser5-RNAPII or P-Ser2-RNAPII nuclear localization (second columns, green signals) detected by PLA. DNA was stained with Hoechst (blue). Scale bar = 2.5 µm. D, In vitro pull-down assays using recombinant AtPRP40b and SE proteins (both full length and the shortened C-terminus variant Δ681–720 were used) and biotinylated CTD peptides in unphosphorylated (UnP) or phosphorylated Ser5 (P-Ser5-CTD) or Ser2 (P-Ser2-CTD) forms. The recombinant proteins were incubated with CTD peptides immobilized on beads. The obtained complexes were washed, eluted, and analyzed by immunoblot. Input represents 1/10 of the protein sample.

  • Additional Information
  • Additional information (application): Suggested blotting conditions: 8% gel, tank blotting, 200 mA/ 1h to nitrocellulose membrane
  • Background
  • Background:

    Serrate RNA effector molecule is required for proper processing of primary miRNAs to miRNA. Also critical for the accumulation of the trans-acting small interfering RNA (ta-siRNA).

  • Product Citations
  • Selected references: Liang et al. (2022). Arabidopsis RBV is a conserved WD40 repeat protein that promotes microRNA biogenesis and ARGONAUTE1 loading. Nat Commun. 2022 Mar 8;13(1):1217. doi: 10.1038/s41467-022-28872-x. PMID: 35260568; PMCID: PMC8904849.
    Li et al. (2023). JANUS, a spliceosome-associated protein, promotes miRNA biogenesis in Arabidopsis. Nucleic Acids Res . 2023 Nov 22:gkad1105. doi: 10.1093/nar/gkad1105.
    Li et al. (2021). In vitro Reconstitution Assays of Arabidopsis 20S Proteasome. Bio-protocol 11(7): e3967. DOI: 10.21769/BioProtoc.3967.
    Li et al. (2020). Apple SERRATE negatively mediates drought resistance by regulating MdMYB88 and MdMYB124 and microRNA biogenesis. Hortic Res. 2020 Jul 1;7:98.doi: 10.1038/s41438-020-0320-6. (ChIP)
    Li et al. (2019). Global co-transcriptional splicing in Arabidopsis and the correlation with splicing regulation in mature RNAs. Mol Plant. 2019 Nov 20. pii: S1674-2052(19)30367-3. doi: 10.1016/j.molp.2019.11.003.
    Wang et al. (2019). The PROTEIN PHOSPHATASE4 Complex Promotes Transcription and Processing of Primary microRNAs in Arabidopsis. Plant Cell. 2019 Feb;31(2):486-501. doi: 10.1105/tpc.18.00556.
    de Francisco Amorim et al. (2018). The U1 snRNP Subunit LUC7 Modulates Plant Development and Stress Responses via Regulation of Alternative Splicing. Plant Cell. 2018 Nov;30(11):2838-2854. doi: 10.1105/tpc.18.00244. Epub 2018 Oct 11.
    Kryovrysanaki et al. (2018). SERRATE, a miRNA biogenesis factor, affects viroid infection in Nicotiana benthamiana and Nicotiana tabacum. Virology. 2018 Dec 29;528:164-175. doi: 10.1016/j.virol.2018.12.011.
    Ma et al. (2018). Arabidopsis Serrate Coordinates Histone Methyltransferases ATXR5/6 and RNA Processing Factor RDR6 to Regulate Transposon Expression. Dev Cell. 2018 Jun 18;45(6):769-784.e6. doi: 10.1016/j.devcel.2018.05.023.
    Wang et al. (2018). SWI2/SNF2 ATPase CHR2 remodels pri-miRNAs via Serrate to impede miRNA production. Nature. 2018 May;557(7706):516-521. doi: 10.1038/s41586-018-0135-x. Epub 2018 May 16.
    Li et al. (2016). Intron Lariat RNA Inhibits MicroRNA Biogenesis by Sequestering the Dicing Complex in Arabidopsis. PLoS Genet. 2016 Nov 21;12(11):e1006422. doi: 10.1371/journal.pgen.1006422. eCollection 2016.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection
  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
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    Number of reviews: (1)
    xiuren zhang | 2020-05-06
    0 | 0
    Excellent antibody for ChIP and western blot.please refer to Dev Cell. 2018 Jun 1845(6):769-784.e6. doi: 10.1016/j.devcel.2018.05.023.Arabidopsis Serrate Coordinates Histone Methyltransferases ATXR5/6 and RNA Processing Factor RDR6 to Regulate Transposon Expression.Ma Z1, Castillo-Gonzlez C1, Wang Z1, Sun D1, Hu X1, Shen X2, Potok ME3, Zhang X4.

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Buy 2 items of this product for 239.00 €/items
Buy 3 items of this product for 217.00 €/items
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