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Synthetic peptide conjugated to KLH derived from within residues 250 to the C-terminus of Human CD82.
(Peptide available as ab71584.)
Our Abpromise guarantee covers the use of ab66400 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 30 kDa).Can be blocked with Human CD82 peptide (ab71584).|
|IHC||Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|IHC-P||1/50 - 1/80. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 22679510|
ab66400 (1:80) staining CD82 in paraffin-embedded human tonsil tissue (left panel) using an automated system (Ventana Discovery). Right hand panel shows negative control (no primary antibody).
Using this protocol there is strong membrane staining of follicular dendritic cells, and the FDC network forming the structure of germinal centres is clearly highlighted by this antibody. There is associated weak staining of smaller cells in the interfollicar areas which may be T cells.
Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Standard Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab66400 (1:80 dilution / 1 hour / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematoxylin and coverslipped in DPX.
For manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
ab66400 (1/50) staining CD82 in paraffin-embedded Mouse salivary gland tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
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