Application
Western blot
Sample
Apis mellifera Tissue lysate - whole (Honey bee whole abdomen lysate)
Gel Running Conditions
Reduced Denaturing (12% Acrylamide)
Loading amount
15 µg
Treatment
Infected with recombinant virus Sindbis-GFP
Specification
Honey bee whole abdomen lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS-T with 5% Milk
Dilution
1/1000
Secondary antibody
Dilution
1/2500
Name
Non-Abcam antibody was used: Rabbit IgG, HRP-linked F(ab')2 fragme
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: 56 kDa
Positive control
None. We expect Ago2 to be present in lysate from infected bees.
Additional data
Additional Notes
We are encouraged by the fact that our dominant band is the same as that found in Drosophila lysate. However, we feel that the band will need to be mass-spec confirmed for future studies.
Image Title: Western Blot using Abcam Drosophila Anti-Argonaute-2 antibody (ab5072) to detect Apis mellifera (honey bee) Argonaute-2 protein (XP_395048.4)
Sample: Apis mellifera Lysate, 48 hrs p.i. with Sindbis-GFP
Additional information: AmelAgo2 (gene: XM_395048.6) and DmelAgo2 are 38% identical at the aa level overall and are 29% identical at the aa level in the target region.
Lane 1: Approximately 15ug honey bee abdomen lysate
Lane 2: Approximately 7.5 ug honey bee abdomen lysate
Lane 3:Approximaterly 1.75 ug honey bee abdomen lysate
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Michelle Flenniken
Verified customer
投稿 Feb 07 2017