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GN | Gnom

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

GN | Gnom in the group Antibodies for Plant/Algal  / Developmental Biology / Signal transduction at Agrisera AB (Antibodies for research) (AS16 3980)

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Product Information

Immunogen

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

Host Rabbit
Clonality Polyclonal
Purity Affinity purrified 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 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 162 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Not reactive in

No confirmed exceptions from predicted reactivity are currently known

Application examples

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

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.

Related products

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.

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