Application
Western blot
Sample
Mouse Tissue lysate - whole (9L, PC12, ms Brain)
Loading amount
10 µg
Specification
9L, PC12, ms Brain
Gel Running Conditions
Reduced Denaturing (Pierce precise protein gel 4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Other product details
Concentration
2 µg/ml
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS[0.1% Tween] +3% MILK
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 West Dura
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 170 kDa Non-specific: various kDa
Negative control
lane 1: 9L (rat brain tumour cell line)
lane 2: PC12 (rat pheochromocytoma cell line)
lane 3: mouse brain whole tissue lysate
lane 2: PC12 (rat pheochromocytoma cell line)
lane 3: mouse brain whole tissue lysate
Additional data
Additional Notes
14 transcripts for this gene of which 9 are protein coding (Ensembl). Therefore may account for some of other bands on the gel though would recommend siRNA to check specificity. Two antibodies AB75014 and AB64158 were tested together on mouse and rat cells. AB75014 reacted well with both species (in agreement with data on the Abcam website).AB64158 only reacted with mouse. (The same lysates were used for each antibody test).
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.
Mr. Antibody Solutions
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投稿 Mar 18 2010