Application
Western blot
Sample
Human Cell lysate - whole cell (Human cerebral organoid)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12.5% gel)
Loading amount
20 µg
Specification
Human cerebral organoid
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: 1% milk in 1X TBS-T
Dilution
1/2500
Secondary antibody
Name
Non-Abcam antibody was used: ECL anti-rat IgG HRP linked whole antibody from GE
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 52-55 kDa
Positive control
Actin in the same lysate
Additional data
Additional Notes
RIPA Lysis buffer used with protease inhibitor ( 20ul for 1ml RIPA)
Lanes from left to right:
1. Organoid control 1 sample
2. Organoid patient 2 sample
3. Organoid patient 3 sample
4. Organoid control 2 sample
5. Organoid patient 4 sample
6. Organoid patient 5 sample
Upper bands are Detyrosinated tubulin and lower bands are Actin
Protein standard ladder used:
Thermo Scientific PageRuler Plus Prestained Protein Ladder (Cat. no. 26619)
Ladder bands from below:
red: 28 KDa
thick blue: 36 KDa
thick blue: ~55 KDa
red: ~72 KDa
thin blue: ~100 KDa
thin blue: ~130 KDa
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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提交于 May 30 2018