Anti-ASY1 | Asynapsis 1

Product no: AS21 4690

AS21 4690  | Clonality:  Polyclonal |  Host:  Rabbit | Reactivity: Hordeum vulgare

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  • Product Info
  • Immunogen: Recombinant ASY1 protein from Hordeum vulgare, UniProt: A0A8I6YI54
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Antigen affinity purified serum, in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl, of sterile or deionized water.
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 1000 (WB)
    Expected | apparent MW: 66.3 kDa
  • Reactivity
  • Confirmed reactivity: Hordeum vulgare
    Predicted reactivity: Arabidopsis thaliana, Hordeum vulgare, Oryza sativa, Triticum aestivum, Zea mays

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-ASY1 antibodies

    0.25 µg/well of total overexpressed Hordeum vulgare ASY1 protein extracted the previous day from Rosetta 2 pLysS cells with BugBuster master mix and Complete EDTA free protease inhibitor, captured with NiNTA, digested at 4°C overnight with ProTEV protease to remove the affinity tag, and run through a second round of NiNTA capture to remove the cleaved affinity tag. Each lane in the gel also includes His-tagged ASY1 protein. Denaturation was performed with LDS buffer, DTT, and Urea at 90°C for 10 min. Samples were separated on 4-12% NuPAGE Bis-Tris SDS-PAGE at 200v for 1h in MOPS buffer with 0.5 ml NuPAGE antioxidant then transferred to PVDF (pore size of 45 µm) at 11 v overnight in NuPAGE transfer buffer with 0.5 ml NUPAGE antioxidant. The blot was fixed in methanol, washed in SDW then blocked with 5 % milk 1h/RT with agitation. The blot was incubated in the primary antibody at a dilution of 1: 5000 for 2h/RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. The blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:25000 in for 1h/RT with agitation. The blot was washed as above and developed for 3 min with Agrisera ECLSuperBright. The exposure time was 37 seconds.

    Courtesy Jamie Orr, The James Hutton Institute, United Kindgdom

  • Background
  • Background: ASY1 (Asynapsis 1) is a protein required for normal meiosis in male and female gametophytes, which plays a crucial role in coordinating the activity of DMC1. Alternative names:  Meiosis-specific protein ASY1.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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