Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (3rd intar larvae)
Loading amount
30 µg
Specification
3rd intar larvae
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% mIlk in TBST
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti Rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Supersignal pico west
Exposure
5 second(s)
Bands
Specific: 58-60 kDa Non-specific: 19, 23, 37, 55(?) kDa
Positive control
Human HFK cells
Negative control
none
Additional data
Additional Notes
Lanes 1 & 2: yW larvae (no transgene). Lane 3: human HFK cells.
Not sure if the extra band below what appears to be the correct size for
Drosophila Tip60 (~ 61 KDa) is an artifact, breakdown or modified form.
I had to let the detection die for about 1.5 hrs before exposing to film.
Next time I will dilute the Tip60 Ab to 1:1,500 or higher depending on sample concentration. Also, another blot blocked with BSA was awful!
Not sure if the extra band below what appears to be the correct size for
Drosophila Tip60 (~ 61 KDa) is an artifact, breakdown or modified form.
I had to let the detection die for about 1.5 hrs before exposing to film.
Next time I will dilute the Tip60 Ab to 1:1,500 or higher depending on sample concentration. Also, another blot blocked with BSA was awful!
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 Nov 22 2011