Application
Western blot
Sample
Rat Tissue lysate - whole (Rat liver tissue)
Loading amount
120 µg
Specification
Rat liver tissue
Treatment
ovariectomy
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
5% milk in TBS-T(0.1%) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 30 minute(s) · Temperature: 4°C · Diluent: 2.5% BSA in TBS-T(0.1%)
Secondary antibody
Name
Non-Abcam antibody was used: Anti-mouse IgG, HRP-linked Antibody
Host species: Horse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Horse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
Pierce ECL West Pico
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 50 kDa Non-specific: 35 kDa
Positive control
Liver tissue lysate was used
Additional data
Additional Notes
The primary antibody could be further diluted to decrease non specific staining. The tissue was homogenised in RIPA buffer containing 1mM PMSF, 1µM aprotinin, 1µg/ml Pepstatin A, 0.5 µg/ml okadaic acid, 1 mM benzamidine HCl and briefly sonicated. The supernatant was used as the total lysate. Proteins were transfered onto nitrocellulose membrane using semi dry transfer.
Lane 1: Liver tissue of control rats
Lane 2: Liver tissue of ovariectomized rats
Lane 1: Liver tissue of control rats
Lane 2: Liver tissue of ovariectomized rats
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.
Mrs. Smitha Koshy
Verified customer
投稿 May 21 2012