Recombinant Anti-CD19 antibody [EPR5906] (ab134114)

Intracellular Flow Cytometry analysis of Raji cells (Human Burkitt's lymphoma B lymphocyte)labelling CD19 with ab134114 at 1/1000 dilution, 1.186µg/ml (red). Cells were fixed with 4% paraformaldehyde, permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/2000. 

Isotype control (black) - Rabbit monoclonal IgG (ab172730) 

Unlabeled control (blue) - Unlabelled cells 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD19 with unpurified ab134114 at a dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunocytochemistry/Immunofluorescence analysis of Raji cells labelling CD19 with purified ab134114 at a dilution of 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

Exposure time:
Lane 1-3: 3 minutes
Lane 4: 30 seconds

Jurkat is a CD19 null cell line according to the paper (PMID:  19147785).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD19 with purified ab134114 at a dilution of 1/500. Heat mediated antigen retrieval was performed using 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.

Negative tissue: Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD19 with purified ab134114 at a dilution of 1/500. Heat mediated antigen retrieval was performed using 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B-cell lymphoma tissue labelling CD19 with unpurified ab134114.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD19 with unpurified ab134114.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B-cell chronic lymphocytic leukaemia tissue labelling CD19 with unpurified ab134114.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue using unpurified ab134114 showing negative staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

IHC image of ab134114 staining CD19 in human tonsil frozen tissue sections, performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab134114 at 1µg/ml for 30 mins at room temperature and detected using an HRP conjugated compact polymer system (Bond™ Polymer Refine Detection). DAB was used as the chromogen. Positive membrane staining was seen in human tonsil. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

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.

IHC image of ab134114 staining CD19 in human heart frozen tissue sections, performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab134114 at 1µg/ml for 30 mins at room temperature and detected using an HRP conjugated compact polymer system (Bond™ Polymer Refine Detection). DAB was used as the chromogen. No staining in human heart (negative tissue) was observed. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

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.