Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD3 antibody [SP7]

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Abreviews
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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Infarcted heart tissue)
Specification
Infarcted heart tissue
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%

Other product details

Dilution
1/200
Incubation time
8 hour(s) and 0 minute(s)

Secondary antibody

Name
Non-Abcam antibody was used: Donkey anti-rabbit Alexa Fluor 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488

Additional data

Additional Notes
The CD3 antigen is expressed on early thymocytes, thymocytes, mature T lymphocytes in peripheral blood and 20-40% of splenic lymphocytes. CD3 antigen is also present in majority of neoplasm of T cell origin, including T-CLL, T-ALL, pleomorphic T-cell lymphomas, and mycosis fungoides. Therefore, CD3 staining is frequently used as an early marker of T-cell differentiation as well as for detecting normal and neoplastic T-cell in tissue sections.

Here, we used infracted rat heart tissue for CD3 staining. Myocardial infarction (MI) has been produced in rat model following ligation of left anterior descending (LAD) coronary artery. Tissue were harvested 6 w following infarct, fixed with Histochoice for 72 hr, paraffin sectioned and slide have been baked prior to CD3 staining. Primary antibody anti-CD3, 1:200 dilutions, incubated overnight at 4 degree centigrade. Image has been taken with a confocal laser scanning microscope. Image (X400 magnification) depicts cells exhibit strong inmmunofluorescence staining for CD3 antigen (green), indicating presence of cells of T-lymphocytes origin in the infarct zone of the heart tissue, counterstained nuclei with DAPI (blue). Note here, CD3 tended to be present in nests of 2-5 cells that were nonuniformly distributed in the infarct zone. In addition, image demonstrated that the CD3 localization is predominantly membrane based and to a certain extent intracytoplasmic. This image implies that T-lymphocytes may play an active and dynamic role in perpetuation of late MI.
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Dr. Mal Niladri

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Submitted Nov 20 2006

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