All lanes : Anti-FOXP3 antibody [mAbcam 450] (ab450) at 1 µg/ml

Lane 1 : HEK293 cells overexpressing human FOXP3
Lane 2 : HEK293 cells overexpressing human FOXP3 with Human FOXP3 peptide (ab16809) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 48 kDa


Exposure time: 45 seconds

Clone mAbcam 450 used in immunocytochemistry on paraffin embedded transfected COS-1 cytospins.

1: FOXP1 transfectant
2: FOXP2 transfectant
3: FOXP3 transfectant
4: FOXP4 transfectant

Strong staining can be seen on the FOXP3 transfectant, with very weak staining on the FOXP1 transfectant and no staining is seen with the other FOXP protein transfectants. This shows that the antibody is largely specific for FOXP3 when used with paraffin embedded cells.

These images were produced with neat supernatant produced with this clone (the stock of this antibody is purified from supernatant).

IHC image of FOXP3 staining in a section of formalin-fixed frozen normal human tonsil* performed on a Leica BOND^TM system using the standard protocol F. The section was then incubated with ab450, 5ug/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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

Clone mAbcam 450 used in immunocytochemistry on transfected COS-1 cytospins fixed with acetone for 10 mins.

1: FOXP1 transfectant
2: FOXP2 transfectant
3: FOXP3 transfectant
4: FOXP4 transfectant

Staining can be seen on the FOXP3 transfectant, but no staining is seen with the other FOXP protein transfectants, confirming the specificity of this antibody in immunocytochemistry.

These images were produced with neat supernatant produced with this clone (the stock of this antibody is purified from supernatant).

IHC image of FOXP3 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BOND^TM 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 20mins. The section was then incubated with ab450, 1ug/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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre