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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
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Name: CD10/CALLA Antibody |
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Clone: 56C6, Mouse anti-CD10/CALLA |
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Supplier: Lab Vision |
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Catalog Number: MS-728-S1 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
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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 |
References: