Immunohistochemistry (Frozen sections) abreview for Anti-eNOS (phospho S1177) antibody

Below Average
Abreviews
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (rat aorta)
Permeabilization
Yes - 0.1% triton X-100
Specification
rat aorta
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Other product details

Incubation time
17 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% goat serum in PBS
Dilution
1/100

Secondary antibody

Dilution
1/1000
Name
Non-Abcam antibody was used: Life technologies Alexa 633 Anti Rabbit IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 647

Additional data

Additional Notes
-The green is autofluorescence with increased brightness and contrast to visualize the extra cellular matrix position. - General expression for endothelial cells was not observed. There were some areas with the red staining but they did not appear to be co-located with cell nuclei. -We have also separately evaluated and submitted an abreview for this antibody using rat aortic endothelial cultured cells, and the antibody seemed to work pretty well using those cells. - Also for the secondary antibody we used Alexa Fluor 633 which was not included in the list. So I just chose the closest one.
Abcam response
Ab184154 is yet to be validated with frozen sections. Reviewing the protocol details we believe, fixing the sections with acetone or methanol would help in getting good staining. The end user has tested this antibody under our Abtrial offer and they are now entitled for a free product in exchange of this review.

Mozhgan Shojaee

Verified customer

Submitted Aug 04 2017

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