Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD20 antibody [EP459Y]

Poor
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spleen)
Specification
Spleen
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Other product details

Dilution
1/100
Incubation time
1 hour(s) and 0 minute(s) · Diluent: Dako

Secondary antibody

Name
Non-Abcam antibody was used: DAKO Rabit Envision HRP
Host species: Rabbit
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase

Additional data

Additional Notes
Immunohistochemical staining with this antibody resulted in negative staining on mouse spleen. For a positive control human tonsil was immunostained and positive.
Additionally cytospins were tested - Raji cells (human B lymphocyte cell line) were positive while A20 cells (mouse B lymphocyte cell line) were negative. The staining indicates that the antibody does not recognize murine CD20.
Abcam response
Thank you for your valuable review. To our knowledge, the antibody has not been tested on mouse samples before. There are several amino acid mis-matches between the immunogen, derived from the human CD20 C-terminus, and the homologous mouse sequence, and this likely accounts for the lack of reactivity with your samples. (The same reviewer also provided data demonstrating positive reactivity with human tonsil).

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Submitted Apr 25 2011

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