WUS | WUSCHEL protein

AS11 1759 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana


product information
Background		WUSCHEL is a transcription factor which plays a central role during early embryogenesis, oogenesis, flowering.
Immunogen		KLH-conjugated peptide derived from Arabidopsis thaliana WUSCHEL protein sequence, UniProt: Q9SB92, TAIR:At2g17950
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Affinity purified serum in PBS, pH 7.4
Format		Liquid
Quantity		50 µg
Reconstitution
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)
Related products		Antibodies to proteins involved in photomorphogenesis Antibodies to DNA/RNA/cell cycle Plant protein extraction buffer Secondary antibodies
Additional information

application information
Recommended dilution		1 : 1000 (WB)
Expected \| apparent MW		33.1 \| kDa
Confirmed reactivity		Arabidopsis thaliana
Predicted reactivity		Arabidopsis thaliana
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references		Dory et al. (2016). Kinase-Associated Phosphoisoform Assay: a novel candidate-based method to detect specific kinase-substrate phosphorylation interactions in vivo. BMC Plant Biol. 2016 Sep 21;16(1):204.

Application information

western blot using anti-WUS antibodies

Plant material: Col-0 wild type (1), pWUS::WUS-GFP (2), expression level comparable to wild type pUbiquitin::mCherry-GR-WUS (3), over-expresses the wuschel protein and also has ectopic expression. Used as a positive control for the westerns. 200mg of 5-7 day old seedling were extracted with 600 μl of 1x SDS-PAGE loading buffer (50mM Tris-HCl pH 6.8, 2% SDS, 10% glycerol, 1% β-MeSH, 12.5mM EDTA, 0.02% bromophenol blue) supplemented with 1% Sigma Protease Inhibitor Cocktail (Cat. Num: P9599). Protein amounts in the extracts were determined by amidoblack. 80 μg (lane 1 and lane 2) or 20 μg (lane 3) of total protein was separated on 10 % SDS-PAGE and blotted for 1.5h to PVDF using semi-dry transfer. Blots were blocked with 4% Milk and 1% BSA in 1x TBST for 1h at room temperature (RT) with agitation. The blot was incubated in the primary antibody at a dilution of 1:1 000 in 1x TBST (0.1% Tween-20) with 1% BSA for 16h at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 3x for 10 min in 1x TBST at RT with agitation. Blot was incubated in secondary antibody diluted to 1:5 000 in 1x TBST for 1h at RT with agitation. The blot was washed as above and developed for 2 min with ECL (Advansta, K-12045-D20) according to the manufacturer's instructions. Exposure time was 120 seconds.

The upper black arrow is mCherry-GRP-linker-WUS. It is a N-terminal fusion product of WUS, while the GFP is a C-terminal linked product (WUS-GFP).

Courtesy of Matyas Medzihradszky, Centre for Organismal Studies, University of Heidelberg, Germany

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