Application
Western blot
Sample
Human Cell lysate - whole cell (PC3 cell line, human prostate carcinoma)
Gel Running Conditions
Reduced Denaturing (NuPAGE 3-8% Tris-Acetate Gel)
Loading amount
38 µg
Treatment
60 ng/ml doxycycline for 72 hours for shRNA induction
Specification
PC3 cell line, human prostate carcinoma
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5 % BSA in TBS-T
Dilution
1/4000
Secondary antibody
Dilution
1/15000
Name
Non-Abcam antibody was used: IRDye 680RD Goat anti-Rabbit IgG (H + L)
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye 680RD
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye 680RD
Detection
Detection method
Odyssey CLx near infrared scanner
Negative control
PC3 with p300 knockdown
Exposure
1 second(s)
Bands
Specific: 300 kDa
Positive control
PC3 cell lysate
Additional data
Additional Notes
Lanes:
1) Ladder
2) PC3 parental cell line
3) PC3 with dox. inducible p300 shRNA, untreated
4) PC3 with dox. inducible p300 shRNA, doxcycline treated
Sample preparation:
Sample Lysis directly in NuPAGE LDS Sample buffer with subesquent sonication Transfer to a PVDF Membrane using the NuPAGE Transfer Buffer in a XCell II Blot Module at 35 V for 2 hours
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. Florian Handle
Verified customer
投稿 Aug 31 2017