GTA MCP | Gene Transfer Agent (GTA) major capsid protein (MCP)
AS08 365 | clonality: polyclonal | host: rabbit | reactivity: Rhodobacterales
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|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||
31.4 | 32 kDa (predicted mature capsid protein of Rhodobacter capsulatus)
|Confirmed reactivity||Rhodobacter capsulatus, Ruegeria pomeroyi DSS-3|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||Mercer and Lang (2014). Identification of a predicted partner-switching system that affects production of the gene transfer agent RcGTA and stationary phase viability in Rhodobacter capsulatus. BMC Microbiol. 2014 Mar 19;14(1):71.|
Roseobacter capsulatus cells, pelleted by centrifugation and resuspended in an equial volume of TE buffer and supernatant sample* were separated on 10% SDS-PAGE and blotted 1h to nitrocellulose. The "+" indicated R. capsulatus SB1003 (GTA positive) and "-" indicated R. capsulatus A1 (GTA capsid protein negative). Blots were blocked in 5% skim milk in TBST followed by incubation with anti-GTA antibodies (AS08 365) at dilution 1: 1000 at 4°C over night. After washes blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Santa Cruz Biotechnology, Santa Cruz, CA) and specific bands were detected with SuperSignal West Femto (Pierce Biotechnology).Exposure time was 30 seconds with CCD camera.
* - supernatant sample was obtained in a following way: cells were removed by two rounds of centrifugation at 17,000 g for 2 min. with sub-sample of the supernatant removed to a new tube. Two volumes of the cells or final culture supernatant were mixed with 1 volume of 3X SDS-PAGE sample buuffer (NEB).
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