All lanes : Anti-Oct-2 antibody [9A2] (ab243153) at 1/1000 dilution

Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 2 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 3 : Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate
Lane 4 : A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate
Lane 5 : Human tonsil tissue lysate
Lane 6 : Mouse spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution

Predicted band size: 51 kDa

The molecular weight observed is consistent with what has been described in the literature (PMID: 3265124).

Exposure times: Lanes 1-3: 3 seconds; Lane 4: 3 minutes; Lane 5: 15 seconds; Lane 6: 44 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Oct-2 with ab243153 at 2.015µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat spleen (PMID: 11280769). The section was incubated with ab243153 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Oct-2 with ab243153 at 2/015µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse spleen (PMID: 11280769). The section was incubated with ab243153 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Oct-2 with ab243153 at 2.015µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on human spleen (PMID: 11280769). The section was incubated with ab243153 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Oct-2 with ab243153 at 2.015µg/ml, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on human tonsil (PMID: 11280769). The section was incubated with ab243153 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Raji (Human Burkitt's lymphoma B lymphocyte) cell line labeling Oct-2 with ab243153 at 1.343µg/ml (red) compared with Rat monclonal IgG (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rat IgG (Alexa Fluor^® 488) (ab150157) at 1/2000 dilution was used as the secondary antibody.