CD90 (Thy-1) Antibody Staining Protocol for Immunohistochemistry

 

Description: The OX-7 antibody reacts with the rat Thy-1 antigen (CD90)1 expressed by hematopoietic stem cells, early myeloid and erythroid cells,4 immature B lymphocytes in the bone marrow and peripheral lymphoid organs, thymocytes, recent thymic emigrants (a subset of CD45RC- peripheral T lymphocytes), neurons, glomerular mesangial cells, endothelium at inflammatory sites, mast cells, and dendritic cells. Rat dendritic epidermal T cells (DEC) are Thy-1-negative, unlike those of the mouse. CD90 is a GPI-anchored membrane glycoprotein of the Ig superfamily which is involved in signal transduction. OX-7 antibody cross-reacts with the mouse Thy-1.1 alloantigen of the AKR/J and PL strains, but not Thy-1.2 found on most mouse strains. In the mouse, CD90 is found on thymocytes, most peripheral T lymphocytes, some intraepithelial T lymphocytes (IEL, DEC), hematopoietic stem cells, and neurons, but not B lymphocytes. In addition, there is evidence that CD90 mediates adhesion of mouse thymocytes to mouse thymic stroma. The OX-7 antibody has also been reported to cross-react with rabbit and guinea pig thymus, brain, and intestine.

 

Primary Antibody

Name: CD90(Thy-1)/CD90.1(Thy-1.1) Antibody

Clone: Mouse anti-Rat

Supplier: BD Pharmingen

Catalog Number: 554895

Dilution: 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60min/room temperature


Antigen Retrieval
Device: Antigen retrieval is not needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Membrane/cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Thymus, bone marrow
Fixation: Acetone fixed frozen sections
Positive Control: Thymus, bone marrow
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

 

References: 

1. Hermans MHA and Opstelten D (1991) In situ visualization of hemopoietic cell subsets and stromal elements in rat and mouse bone marrow by immunostaining of frozen sections. J Histochem Cytochem. 39 (12):1627-34.