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Anti-N-YFP | N-terminal of YFP
Product no: AS11 1776
AS11 1776 | Clonality: Polyclonal | Host: Rabbit | Reactivity:N-terminal of YFP
329
€
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- Product Info
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Immunogen: KLH-conjugated synthetic peptide derived from N-terminal of YFP protein.
Host: Rabbit Clonality: Polyclonal Purity: Serum Format: Lyophilized Quantity: 50 µl 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 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 : 2000 (WB) - Reactivity
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Confirmed reactivity: N-YFP tagged proteins from Arabidopsis thaliana, Escherichia coli, Nicotiana tabacum Not reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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Application example
Arabidopsis thsliana leaf lysate which is co-expressing test protein 1 with nYFP tags (~60 kDa) and test protein 2 with cYFP-tag (~40 kDa). In each case only one band is visible demonstrating that the antibodies do not cross-react. Sample preparation and immunoblot analysis were carried out as described in Karnik et al., Plant Cell 2015, March 2015 vol. 27 no. 3 675-694. For immunoblot analysis of plant tissues,leaves were excised and flash frozen in liquid N2. Frozen tissue was ground in equal volumes (w/v) of homogenization buffer containing 500 mM sucrose, 10% glycerol, 20 mM EDTA,20mMEGTA,ProteaseInhibitor(Roche),10mMascorbicacid,5mM DTT, and 50 mM Tris-HCl, pH 7.4, and centrifuged at 13,000g and 4°C for 30 min to pellet debris. Supernatant was diluted 1:1 in 23 Laemmli buffer containing 2.5% 2-mercaptoethanol, heated to 95°C for 10 min, and separated by SDS-PAGE on a 12% Acrylamide gel. Ponceau S-stained Rubisco bands were used as loading standards for plant samples. Membrane type: Cellulose Nitrate (GE Healthcare) Blocking reagent: GE Healthcare Wash buffer: Tris Buffered Saline, 0.5% Tween Exposure time: 10 – 20 seconds.
Courtesy of Dr. Rucha Karnik, University of Glasgow, UK
- Additional Information
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Additional information (application): YFP molecular weight is 27 kDa, however detected band is going to have MW increased by a fused protein - Background
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Background: YFP (Yellow Fluorescent Protein) is a genetic mutant of green fluorescent protein (GFP). YFP has an excitation peak at 514 nm and emission peak at 527 nm. - Product Citations
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Selected references: Li et al. (2021) Two ubiquitin-associated ER proteins interact with COPT copper transporters and modulate their accumulation, Plant Physiology, 2021;, kiab381, https://doi.org/10.1093/plphys/kiab381
Lung et al. (2021) Oxylipin signaling in salt-stressed soybean is modulated by ligand-dependent interaction of Class II acyl-CoA-binding proteins with lipoxygenase. Plant Cell. 2021 Dec 17:koab306. doi: 10.1093/plcell/koab306. Epub ahead of print. PMID: 34919703.
Schultz-Larsen et al. (2018). The AMSH3 ESCRT-III-Associated Deubiquitinase Is Essential for Plant Immunity. Cell Rep. 2018 Nov 27;25(9):2329-2338.e5. doi: 10.1016/j.celrep.2018.11.011. - Protocols
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Antibody protocols
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