Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Denaturing (8)
Sample
Xenopus laevis Tissue lysate - whole (whole embryo)
Specification
whole embryo
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Other product details
Incubation time
15 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA (1x TBST)
Dilution
1/100
Secondary antibody
Name
Non-Abcam antibody was used: IR Dye 680 LT Conjugated Donkey Anti-Goat IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 680LT
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 680LT
Dilution
1/10000
Detection
Detection method
IR (Licor System)
Bands
Specific: - kDa Non-specific: too many kDa
Exposure
2 minute(s) and 0 second(s)
Positive control
overexpressed Xenopus foxa2 protein (injection of mRNA into 2-cell stage Xenopus embryos, phenotype detectable)
Additional data
Additional Notes
tested on whole Xenopus embryo lysate (neurula stage, foxa2 highly abounded)
tested on overexpressed Xenopus foxa2 protein (foxa2-mRNA injected in early stage Xenopus embryos)
blocking for 1 or 2 h
AB tested in different concentrations (from 1 ´g to 1:50 dilution)
AB incubation in 5% BSA (in 1x TBST) or 1x TBST w/o BSA for 1h at rt or 4¯C o/n
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.
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投稿 Apr 10 2015