product information

background
 

Gene Transfer Agents (GTAs) are bacteriophage-like particles which function as a means to package and transfer genomic DNA between bacterial cells.GTA-mediated gene transfer might be an important phenomenon in natural microbial communities.

immunogen
 

KLH-conjugated conserved peptide sequence found in the Gene Transfer Agent (GTA) major capsid protein (MCP) encoded in Bacteria within the Rhodobacterales order of the class alpha-Proteobacteria including Rhodobacter sphaeroidesQ3J3K4

 

antibody format
 

rabbit;

polyclonal;

serum;

lyophilized

quantity
 

200 µl

- for reconstitution add 200 µl of sterile distilled 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)

related products
 

to be added when available

additional information
 

to be added when available

application information

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

predicted reactivity
 

Rhodobacterales

not reactive in
 

no confirmed exceptions from predicted reactivity known in the moment

additional information
 

to be added when available

selected references
 

Yunyun et al. (2010). High diversity of Rhodobacteriales in the subarctic North Atlantic Ocean and gene transfer agent protein expression in isolated strains. Aquatic Microbial Ecology.59:283-293.

application example

 

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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