Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Cor 1 cells - unstimulated)
Specification
Cor 1 cells - unstimulated
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% TWEEN 20
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Other product details
Dilution
1/100
Incubation time
2 hour(s) and 0 minute(s) · Diluent: TBS/BSA/TWEEN 20/azide
Secondary antibody
Name
Non-Abcam antibody was used: anti Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Dilution
1/1000
Additional data
Additional Notes
My image appears to be very similar to the IF HeLa cell image shown on the Abcam data sheet: a strong, diffuse cytoplasmic positivity, although my results indicate a more punctate positivity.
I am not familiar with FAK expression profiles yet.
Fixation: equal vol of 4% PFA was added to the TCS, for 10 mins, removed, then 4% PFA was applied, for 10mins.
NB: this is not generally required but, if one is wishing to preserve nerve cell growth cone morphology, for example, it has been shown that this more "gentle" approach is less disruptive ( advice from Professor Britta Eickholt, MRC Centre for Developmental Neurobiolgy, Kings College London)
After washing in PBS, wells were aspirated and TBS/BSA/azide/0.1% Tween20 was added, for 20mins.
Primary Ab was diluted in same buffer.
NB: I prefer TRIS-based buffers although I have not noticed any difference between PBS and TRIS buffers, for IF applications.
I am not familiar with FAK expression profiles yet.
Fixation: equal vol of 4% PFA was added to the TCS, for 10 mins, removed, then 4% PFA was applied, for 10mins.
NB: this is not generally required but, if one is wishing to preserve nerve cell growth cone morphology, for example, it has been shown that this more "gentle" approach is less disruptive ( advice from Professor Britta Eickholt, MRC Centre for Developmental Neurobiolgy, Kings College London)
After washing in PBS, wells were aspirated and TBS/BSA/azide/0.1% Tween20 was added, for 20mins.
Primary Ab was diluted in same buffer.
NB: I prefer TRIS-based buffers although I have not noticed any difference between PBS and TRIS buffers, for IF applications.
Abcam response
This negative Abreview is currently being investigated by Abcam Technical Support. Please check here for an update response from Abcam.
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. Carl Hobbs
Verified customer
投稿 Jul 19 2010