Immunohistochemical analysis of paraffin-embedded human thymus tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in human thymus (PMID: 16380964) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for CTLA4 using ab237712 at 0.25 μg/ml in immunohistochemical analysis.

Incubate with primary antibody for 75 minutes at room temperature.

Anti-CD4 antibody [EPR6855] (ab133616)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with ab133616 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

IHC image of HLA-DR [TAL 1B5] staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20181, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunohistochemical analysis of paraffin-embedded human Hodgkin’s lymphoma tissue labeling CCR6 with ab227036 at 1/1000 dilution, followed by the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on human Hodgkin’s lymphoma (PMID: 12514792) is observed. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with purified ab133616 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.