GN | Gnom

Product no: AS16 3980

AS16 3980 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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  • Data sheet
  •  
  • Product Info
  • Immunogen:

    KLH-conjugated peptide derived from Arabidopsis thaliana GN protein sequence, UniProt: Q42510, TAIR: At1g13980

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 g
    Reconstitution: For reconstitution add 50 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; 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: Western blot (WB)
    Recommended dilution: 1 : 1000 (WB)
    Expected | apparent MW: 162 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-GN antibodies

    Samples: M - Marker, A - Arabidopsis thaliana wild type, B - gnom R5 mutant, C gn:GNOM-GFP/gn:GNOM-3xHA The cell lysate was prepared from 10 A. thaliana seedlings (8-9d old). Seedlings were frozen in liq N2, then lysed with tissue lyzer, and 100 µl extraction buffer (100mM NaH2PO4 10mM Tris/HCl pH 8.0, 8M Harnstoff) was added. Total protein was denatured with 4x Laemmli buffer at 72ºC for 5min and separated on a 4-20% Mini Protean precast gel from BioRad and blotted for 4.5h to nitrocellulose using wet transfer. Blots were blocked with 5%NFDM for 1h at RT with agitation. The blot was incubated with primary antibody at a dilution of 1:1000 O/N at 4ºC in 1% NFDM in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once 15min and 3x 5min in TBS-T at RT with agitation. The blot was incubated using anti-rabbit IgG (HRP conjugated) from Agrisera (AS09 602) diluted 1:25 000 in 1% NFDM in TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5min with Agrisera ECLSuperBrigth Exposure time was 5 sec.
    The cell lysate (GNOM-GFP/GNOM-3xHA) was prepared by grinding 1g seedlings(5-6 days old) in 1ml lysis buffer (50mM Tris-HCl pH7.5, 150mM Nacl+2mM EDTA+1% Tx 100 + Protease inhibitors). Total protein was denatured with 4x Laemmli buffer at 95C for 5min.

    Courtesy of Dr. Antje Feller, University of Tübingen, Germany

  • Additional Information
  • Additional information (application):

    Nitrocellulose membrane with pore size 0.45 µm is recommended as well as TBS-T as antibody incubation buffer. 

    This antibody is also recognzing recombinant GN fused to GFP.

  • Background
  • Background: GN (Gnom) activates the ARF proteins by exchanging bound GDP for free GTP and plays a role in vesicular protein sorting. Acts as the major regulator of endosomal vesicle trafficking but is also involved in the endocytosis process. GN is required for correct cell wall organization leading to normal cell adhesion during seedling development amd plays also an essential role in hydrotropism of seedling roots.
    Alternative names: Pattern formation protein EMB30, Protein EMBRYO DEFECTIVE 30, Protein MIZU-KUSSEI2, Protein VASCULAR NETWORK 7
  • Product Citations
  • Selected references: To be added when avaibale, antibody released in December 2020.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection
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