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Dendritic Cells Antibody Staining Protocol for Immunohistochemistry
Description: Specificity: Anti-Mouse Dendritic (clone 33D1) recognizes a mouse cell surface antigen on dendritic cells. Antigen Distribution: The antibody is cytotoxic and does not appear to be directed against la or immune response associated antigen. The antigen is present only on dendritic cells with an estimated frequency of 10,000-15,000 sites per cell.
Primary Antibody
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Name: Dendritic Cells Antibody |
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Clone: 33D1, Rat anti-Mouse |
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Supplier: Leino Technologies |
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Catalog Number: D104 |
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Dilution: 1:500 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: 60 minutes/room temperature |
Antigen Retrieval
| Device: N/A |
| 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 |
| Images: Search image |
Additional Information:
| Tissue Type: Mouse spleen |
| Fixation: Acetone or formalin fixed frozen sections |
| Positive Control: Mouse spleen |
| 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:
Not tested on paraffin sections |
References: