Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (LLC-PK1 cell line)
Permeabilization
Yes - 0.5% Triton-X 100
Specification
LLC-PK1 cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: R.T°C
Fixative
Paraformaldehyde
Other product details
Incubation time
0 minute(s) · Temperature: 4°C · Diluent: Blocking buffer
Dilution
1/200
Secondary antibody
Dilution
1/250
Name
Non-Abcam antibody was used: Donkey Anti Mouse Alexa 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
Experimental Steps
Rinse - 1x PBS
Prefix - 4% PFA in PBS - 30 seconds
Permeabilize - 0.5% Triton X in PBS - 5 minutes
Fix - 4% PFA in PBS - 20 minutes
Block - 0.1% BSA in PBS - 1 hour - R.T
Primary Antibody Mre11 - 1:200 in blocking buffer - 0/N at 4°C
Secondary Antibody - Donkey anti Mouse- 1:250 – 1 hour at 37°C
DAPI - 1:10000 – 10 minutes
The experiment was also done using a methanol fixation step following by the same blocking and antibody steps. Both methods are represented in the picture attached.
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 Feb 10 2016