Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibrosarcoma (HT1080))
Specification
fibrosarcoma (HT1080)
Fixative
Methanol
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1%
Other product details
Dilution
1/500
Incubation time
24 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: Alexa Fluor 488-donkey anti-mouse
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
Cells were treated with 0.1-0.2ug/mL colcemid for 45-60 minutes, then swollen in hypotonic buffer for 8 minutes and centrifuged onto glass slides. Cells were blocked in 1X PBS + 1% BSA + 0.5% Triton X-100 (blocking buffer) for 30 minutes at room temperature. Antibodies were diluted 1:300-1:500 in blocking buffer and incubated overnight at 4 degrees C. Antibodies (ab5093) were detected with Alexa Fluor 488-donkey anti-mouse (Molecular Probes/Invitrogen) for 1-2 hours at room temperature.
This image was kindly provided by Kaitlin Stimpson and Dr. Beth Sullivan, Duke University.
This image was kindly provided by Kaitlin Stimpson and Dr. Beth Sullivan, Duke University.
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.
Dr. Beth Sullivan
Verified customer
投稿 Jan 17 2007