CD10 Antibody Staining Protocol for Immunohistochemistry


Description: CD10, also known as Common Acute Lymphocytic Leukemia Antigen (CALLA), is a cell surface enzyme with neutral metalloendopeptidase activity which inactivates a variety of biologically active peptides. CD10 is expressed on the cells of lymphoblastic, Burkitt’s, and follicular germinal center lymphomas, and on cells from patients with chronic myelocytic leukemia (CML). It is also expressed on the surface of normal early lymphoid progenitor cells, immature B cells within adult bone marrow and germinal center B cells within lymphoid tissue. CD10 is also present on breast myoepithelial cells, bile canaliculi, fibroblasts, with especially high expression on the brush border of kidney and gut epithelial cells.


Primary Antibody

Name: CD10/CALLA Antibody

Clone: 56C6, Mouse anti-CD10/CALLA

Supplier: Lab Vision

Catalog Number: MS-728-S1

Dilution: 1:50 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane
Images: Search image

Additional Information:
Tissue Type: Small intestine, kidney, tonsil, thymus, bone marrow, breast
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Small intestine, kidney, tonsil, thymus, bone marrow, breast
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


1. There is no need to perform antigen retrieval for acetone fixed frozen sections

2. CD10 (Clone 56C6, Novocastra, Cat# NCL-CD10-270) also works well at 1:50 dilution