Application
Immunocytochemistry
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% in
Other product details
Dilution
1/200
Incubation time
16 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: Alexa 568 Anti-Rabbit
Host species: Goat
Clonality: Polyclonal
Host species: Goat
Clonality: Polyclonal
Additional data
Additional Notes
HeLa cells, fixed for 10mins in fresh 3.7% paraformaldehyde, permeabilised with PBS + 0.1% Triton X 100, Blocked with 1% goat serum in Abdil.
Primary antibody (in Abdil) incubated OVERNIGHT (1 hour didn't work) at room temp.
Other details above.
Cells co-stained with mouse anti-tubulin (secondary is Anti-Mouse Alexa 488) and ACA (Human anti centromere marker, secondary is Anti-Human Cy5).
Cells also stained with DAPI.
Figure shows projected images of dividing cells (prophase, prometaphase, metaphase, anaphase, telophase). Top panels show ab12183, DNA and microtubule stains. Lower panels are the same cells, showing a subset of optical sections at higher magnification. ACA (geen) stains centromeres and ab12183 (red) stains the inner centromere in prophase, prometaphase, and metaphase. The ab12183 channel has been linerally scaled to reduce the background staining associated with this antibody.
Primary antibody (in Abdil) incubated OVERNIGHT (1 hour didn't work) at room temp.
Other details above.
Cells co-stained with mouse anti-tubulin (secondary is Anti-Mouse Alexa 488) and ACA (Human anti centromere marker, secondary is Anti-Human Cy5).
Cells also stained with DAPI.
Figure shows projected images of dividing cells (prophase, prometaphase, metaphase, anaphase, telophase). Top panels show ab12183, DNA and microtubule stains. Lower panels are the same cells, showing a subset of optical sections at higher magnification. ACA (geen) stains centromeres and ab12183 (red) stains the inner centromere in prophase, prometaphase, and metaphase. The ab12183 channel has been linerally scaled to reduce the background staining associated with this antibody.
Abcam response
Thank you for your excellent review and for submitting the beautiful images. Your results clearly show that this antibody works nicely for ICC. However, as you have pointed out - in order to get the signal it is very important to incubate the cells with ab12183 for overnight (not for 1 hr). We believe that this information will be very useful for our future customers.
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. William Moore
Verified customer
投稿 Nov 30 2005