N-YFP | N-terminal of YFP

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AS11 1776 | Clonality: Polyclonal | Host: Rabbit | Reactivity:N-terminal of YFP

Product Information

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 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 : 2000 (WB)

Reactivity

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

Application example

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

YFP molecular weight is 27 kDa, however detected band is going to have MW increased by a fused protein.

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Secondary antibodies

Background

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

Selected references 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.