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Anti-Patatin
AS12 1842 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Solanum tuberosum
- Product Info
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Immunogen: KLH-conjugated synthetic peptide derived from a C-terminal part of 36 known isoforms of patatin from Solanum tuberosum including: Q3YJS9, Q3YJT0, Q42502, Q3YJT2
Host: Rabbit Clonality: Polyclonal Purity: Serum Format: Lyophilized Quantity: 100 µl Reconstitution: For reconstitution add 100 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: Immunoprecipitation (IP), Immunolocalization (IL), Western blot (WB) Recommended dilution: 1 : 100 (IL), 1 : 2000 (WB) Expected | apparent MW: 40-42 kDa
- Reactivity
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Confirmed reactivity: Solanum tuberosum
Predicted reactivity: Solanum tuberosum Not reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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application example
75 µg of total protein from Solanum tuberosum v. Nicola extracted with 20 mM TRIS PH-8.5, 10 mM thiourea,10mM CaCl2, 5mM DTT, 1 mM PMSF, 1%PVPP were separated on 4-20 % SDS-PAGE and blotted 1.5h to PVDF. Blots were blocked with 5% milk powder in T-TBS (2.5gr of powder in 50 ml T-TBS) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 for overnight at 4°C with agitation. The antibody solution was decanted and the blot was washed 6 times for 5 min. in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:10 000 in blocking solution for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 30 seconds.Courtesy of Dr. Paula Teper-Bamnolker and Dr. Dani Eshel, The Volcani Center, Israel
Proteins were extracted from tuber flesh of Russet Burbank with 0.1 M Tris HCl (pH=8.0), 5% sucrose (m/v), 2% (m/v) SDS, protease inhibitors (PMSF 1mM). Samples were heated 95°C 5 min, and 1 μg of total protein was resolved in 12% SDS PAGE and blotted to PVDF membrane for 1h-1.5h. Blocking with a 4% skimmed milk in T-TBS (1.5h). Primary antibodies were applied overnight +4°C in dilution 1:2000. After washing with T-TBS 2-3 times, membrane was incubated with secondary antibodies (Goat Anti-Rabbit HRP conjugate) 1:10 000 for 1 hour at RT. The blot was developed with ECL (Clarity Western ECL Substrate, BioRad, 170-5060) for 5 – 10 minutes. Exposure time – 0.4 second.
Courtesy of Msc. Iauhenia Isayenka, University of Sherbrooke, Canada - Background
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Background: Patatin is the most abundant protein in potato (Solanum tuberosum L.) tubers, about 60 % of a total protein. It serves as a storage protein. Mature tuber patatin variants are dimers of 40-42 kDa subunits without disulphide bridges.
- Protocols
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Agrisera Western Blot protocol and video tutorials
Imunolocalization
Can be done according to the protocol described by Paciorek et al. (2006). - Reviews:
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Accessories
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