Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HEF1 / NEDD-9 antibody [2G9]

Inconclusive
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tumour: Pagets disease of the nipple)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Tumour: Pagets disease of the nipple
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Formaldehyde

Other product details

Incubation time
2 hour(s) and 0 minute(s) · Diluent: TBS/BSA/azide
Dilution
1/200

Secondary antibody

Dilution
1/200
Name
Non-Abcam antibody was used: anti mouse Igs
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin

Additional data

Additional Notes
I used a standard stABCpx-DAB detection system. I can offer no justification for using this tissue as a "control" tissue other than to explain that I have another Abcam anti Hef1 antibody, ab37161, which I titrated on the same tissue using serial sections and I achieved different results. With ab18056 I noted a slight, seemingly non-specific cytoplasmic positivity, an occassional nucleus that was positive but mostly this appeared to be at the rim of the nuclei ( tumour cells only). Please ignore the darker brown extracellular, globular positivity: this appears to be lipofuscin, which is somehow enhanced by application of this antibody, in my opinion. Please compare and contrast the images submitted for this Ab on human , rat and mouse and those for ab37161. NB: This Ab reagent may not be suitable for paraffin wax sections.
Abcam response
Thank you for your honest and thoughtful feedback. We have yet to test this antibody in IHC-P, but your data suggest it may be unsuitable for this application.

Mr. Carl Hobbs

Verified customer

Submitted Jan 05 2009

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