Application
Western blot
Sample
Zebrafish Tissue lysate - whole (whole embryo)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10 µg
Treatment
25uM tamoxifen
Specification
whole embryo
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBST 2%BSA
Dilution
1/10000
Secondary antibody
Dilution
1/1000
Name
Non-Abcam antibody was used: donkey anti-Rabbit IgG (H+L) Cross Adsorbed Second
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 25 kDa Non-specific: 10, 12 kDa
Positive control
human HEK293T cells untreated and treated with doxorubinin (lanes
Additional data
Additional Notes
ABTRIAL-EBLLK
We have tried additional conditions: dilution 1:10000, buffer without BSA, dilution 1:1000 (PBST 2%BSA)
We could not find the conditions for this antibody to give reproducible results for zebrafish.
For human samples works well.
Lane 1. HEK297 untreated
Lane 2. HEK297 treated doxorubicin
Lane 3. Zebrafish whole body lysate untreated
Lane 4. Zebrafish whole body lysate treated tamoxifen
Lane 5. Zebrafish whole body lysate treated tamoxifen and cisplatin
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 Jul 28 2015