Anti-GUN1 | Pentatricopeptide repeat-containing protein GUN1, chloroplastic
AS23 4931 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
- Product Info
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Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana GUN1 protein sequence, UniProt: Q9SIC9 TAIR: AT2G31400 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: 500 - 1 : 1000 (WB) Expected | apparent MW: 101.6 | 97.2 kDa (due to N-terminal processing) - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Arabidopsis thaliana Not reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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Samples:
1. 50 ug of 36-h-old WT seedlings (Arabidopsis thaliana)
2. 50 ug of 36-h-old gun1 mutant (Arabidopsis thaliana)
3 and 4: 50 ug of two 36-h-old independent overexpression lines with GFP tag in gun1 background (Arabidopsis thaliana)50 µg/well of total protein extracted freshly from 36-h-old Arabidopsis thaliana seedling. Exact buffer components were: 0.0625 M Tris-HCl (pH 6.8), 1% (w/v) SDS, 10% (v/v) glycerol, 0.01% (v/v) 2-mercaptoethanol. and denatured with exact buffer components at 95 °C/5min. Samples were separated on 10% SDS-PAGE at room temperature (RT) and blotted for 1.5 h to Immune-Blot PVDF ( Bio-Rad (0.2 um)) using: wet in the cold. Blot was blocked with 5 % milk for: 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 ON/4°C with agitation. Then washed once for 20 min and 4 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 10 000 for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent. Exposure time was 5 seconds.
Note regarding background bands: the aim of the protocol which was used above was to confirm that GUN1 protein is detected. Background signal can be decreased by adjustment of Western blot protocol: primary antibody incubation at 1: 1000 1h/RT. - Background
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Background: GUN1 (pentatricopeptide repeat-containing protein GUN1) is Involved in nuclear gene expression via the retrograde plastid-to-nucleus signaling.
Alternative names: entatricopeptide repeat-containing protein At2g31400, Protein GENOMES UNCOUPLED 1. - Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts
Agrisera Educational Poster Collection - Reviews:
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