All lanes : Anti-FOXP3 antibody [221D] (ab253297) at 1.514 µg/ml

Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a GFP-myc tag, whole cell lysate
Lane 2 : HEK-293T transfected with FOXP3 (WT) expression vector containing a GFP-myc tag, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 47 kDa
Observed band size: 73 kDa why is the actual band size different from the predicted?


Exposure time: 3 seconds

The molecular weight observed was in consistant with the literature (PMID: 17586580).

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling FOXP3 with ab253297 at 1/1000 dilution (1.514 µg/ml) followed by a ready to use secondary. Nuclear staining on the human tonsil. The section was incubated with ab253297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling FOXP3 with ab253297 at 1/1000 dilution (1.514 µg/ml) followed by a ready to use secondary. Nuclear staining on the stroma cells in the human colon. The section was incubated with ab253297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue labeling FOXP3 with ab253297 at 1/1000 dilution (1.514 µg/ml) followed by a ready to use secondary. Nuclear staining on the stroma cells in the human stomach carcinoma. The section was incubated with ab253297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling FOXP3 with ab253297 at 1/1000 dilution (1.514 µg/ml) followed by a ready to use secondary. Nuclear staining on mouse spleen tissue. The section was incubated with ab253297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling FOXP3 with ab253297 at 1/1000 dilution (1.514 µg/ml) followed by a ready to use secondary. Nuclear staining on mouse thymus tissue. The section was incubated with ab253297 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.