Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling CD103 with ab224202 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the lymphocytes in the rat colon (PMID: 23355207) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD103 with ab224202 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the lymphocytes in the mouse spleen (PMID: 23355207) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling CD103 with ab224202 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the lymphocytes in the human lung carcinoma (PMID: 25725111). The section was incubated with ab224202 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD103 with ab224202 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the lymphocytes in the human colon (PMID: 16775489, 30405182, 23355207). The section was incubated with ab224202 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat colon tissue labeling CD103 with ab224202 at 1/100 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on the lymphocytes in the rat colon (PMID: 16775489) is observed. Counterstained with DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse colon tissue labeling CD103 with ab224202 at 1/100 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on the lymphocytes in the mouse colon (PMID: 16775489) is observed. Counterstained with DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

All lanes : Anti-CD103 antibody [EPR22590-27] (ab224202) at 1/1000 dilution

Lane 1 : Mouse stomach cell lysate
Lane 2 : Mouse spleen cell lysate
Lane 3 : Mouse thymus cell lysate
Lane 4 : Rat stomach cell lysate
Lane 5 : Rat spleen cell lysate
Lane 6 : Rat thymus cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 130 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:26921343; PMID:29021139).

Blocking/Dilution buffer: 5% NFDM/TBST.

Anti-CD103 antibody [EPR22590-27] (ab224202) at 1/1000 dilution + His-tagged human CD103 recombinant protein fragment (aa676-965), 20ng

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 130 kDa


Exposure time: 6 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.