Anti-CD19 antibody [SP291] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Intracellular Flow Cytometry analysis of Raji (human Burkitt's lymphoma) labeling CD19 with purified ab227688 at 1/20 dilution (7.5μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotypecontrol - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human thymus tissue staining CD208 with ab324381 at a 1 : 100 (4.94 ug/ml) dilution, ab281573 anti-LAMP3/CD208 used at 1 : 500 (0.938 ug/ml) dilution and ab237772 anti-CD19 used at a 1 : 5000 (0.21 ug/ml) dilution.

Panel A : merged staining of anti-LAMP3/CD208 (green; Opal™520), anti-LAMP3/CD208 (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on human thymus.
Panel B : anti-LAMP3/CD208 staining dendritic cells in human thymus.
Panel C : anti-LAMP3/CD208 staining dendritic cells in human thymus.
Panel D : anti-CD19 staining B lymphocytes of human thymus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324381, ab271053, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining CD208 with ab324381 at a 1 : 100 (4.94 ug/ml) dilution, ab281573 anti-LAMP3/CD208 used at 1 : 500 (0.938 ug/ml) dilution and ab237772 anti-CD19 used at a 1 : 5000 (0.21 ug/ml) dilution.

Panel A : merged staining of anti-LAMP3/CD208 (green; Opal™520), anti-LAMP3/CD208 (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on human tonsil.
Panel B : anti-LAMP3/CD208 staining dendritic cells in human tonsil.
Panel C : anti-LAMP3/CD208 staining dendritic cells in human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324381, ab271053, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

Panel A : merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen.
Panel B : anti-CD8 alpha stained on cytotoxic T cells.
Panel C : anti-CD4 stained on T helper cells.
Panel D : anti-CD19 stained on B cells.

Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of ab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

Panel A : merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B : anti-CD8 alpha stained on cytotoxic T cells.
Panel C : anti-CD4 stained on T helper cells.
Panel D : anti-CD19 stained on B cells.

Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of ab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This image was generated using ab227688, the same clone, but with a different buffer formulation. Panel A : merged staining of anti-CD68 (magenta; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human spleen. Secondary antibody was Opal Polymer HRP Ms + Rb, and couterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Formalin-fixed, paraffin-embedded human thymus tissue stained for CD19 using ab227688 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Formalin-fixed, paraffin-embedded human spleen tissue stained for CD19 using ab227688 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Intracellular Flow Cytometry analysis of Ramos (human Burkitt's lymphoma cell line) cells, labeling CD19 with ab227688 at 1/400 dilution (green) compared to a Rabbit IgG negative control (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Formalin-fixed, paraffin-embedded human B-Cell lymphoma tissue stained for CD19 using ab227688 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining TCF7 with ab315390 at a 1 : 2000 (0.26 ug/ml) dilution, ab237707 anti-CD3E used at 1 : 500 (1.01 ug/ml) dilution and ab237772 anti-CD19 used at a 1 : 5000 (0.21 ug/ml) dilution.

Panel A : merged staining of anti-TCF7 (magenta; Opal™690), anti-CD3E (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B : anti-TCF7 staining T lymphocytes of human tonsil.
Panel C : anti-CD3E staining T lymphocytes of human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order of ab315390, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This data was developed using ab227688, the same antibody clone in a different buffer formulation

The BOND™ Polymer Refine Detection System on the BOND RX Automated Stainer (DS9800, Leica Biosystems, Wetzlar, Germany) was used for immunohistochemical labelling of formalin-fixed, paraffin-embedded (FFPE) tissues. The protocol includes a peroxide block, post-primary reagent, polymer detection, DAB chromogen, and haematoxylin counterstain, all automated to minimize variability. Antigen retrieval was performed by heat-induced epitope retrieval (HIER) for 20 minutes in Tris-EDTA buffer (BOND Epitope Retrieval Solution 2, AR9640, Leica Biosystems, Wetzlar, Germany). For mouse and rabbit antibodies, the sequence comprised peroxide block (10 min), primary antibody incubation (30 min), post-primary (10 min), polymer (10 min), DAB (10 min), and haematoxylin (8 min). For rat antibodies, an anti-rat step (1 : 300 dilution, Vector Laboratories, California, USA; 20 min) was included prior to polymer application (15 min).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining CD21 with ab315160 at a 1 : 50 (10.56 ug/ml) dilution, ab237772 anti-CD19 used at 1 : 5000 (0.21 ug/ml) dilution and ab237707 anti-CD3E used at a 1 : 500 (1.01 ug/ml) dilution.

Panel A : merged staining of anti-CD21 (green; Opal™690), anti-CD3E (grey; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B : anti-CD21 staining mature B lymphocytes and T-lymphocytes of human tonsil.
Panel C : anti-CD19 staining B lymphocytes of human tonsil.
Panel D : anti-CD3E staining T lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order of ab315160, ab237772, and ab237707 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for CD19 using ab227688 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, and sodium azide (ab227688).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Formalin-fixed, paraffin-embedded human colon tissue stained for CD19 using ab227688 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227688).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining TCF7 with ab315392 at a 1 : 2000 (0.26 ug/ml) dilution, ab237707 anti-CD3E used at 1 : 500 (1.01 ug/ml) dilution and ab237772 anti-CD19 used at a 1 : 5000 (0.21 ug/ml) dilution.

Panel A : merged staining of anti-TCF7 (magenta; Opal™690), anti-CD3E (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B : anti-TCF7 staining T lymphocytes of human tonsil.
Panel C : anti-CD3E staining T lymphocytes of human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order of ab315392, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver cancer labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 dilution(D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human liver cancer.

Panel B : anti-CD208 staining dendritic cells in human liver cancer.

Panel C : anti-CD3E staining T lymphocytes in human liver cancer.

Panel D : anti-CD19 staining B lymphocytes of human liver cancer.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 dilution(D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human tonsil.
Panel B : anti-CD208 staining dendritic cells in human tonsil.
Panel C : anti-CD3E staining T lymphocytes in human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 dilution(D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human colon.

Panel B : anti-CD208 staining dendritic cells in human colon.

Panel C : anti-CD3E staining T lymphocytes in human colon.

Panel D : anti-CD19 staining B lymphocytes of human colon.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 dilution(D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human colon cancer.

Panel B : anti-CD208 staining dendritic cells in human colon cancer.

Panel C : anti-CD3E staining T lymphocytes in human colon cancer.

Panel D : anti-CD19 staining B lymphocytes of human colon cancer.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Panel A : merged staining of anti-CD31 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human spleen. Secondary antibody was Opal Polymer HRP Ms + Rb, nuclear counterstain was DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD31 stained on endothelial cells with ab207090 at 1/500 dilution The section was incubated in three rounds of staining : in the order of ab207090 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins"

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This data was developed using ab16669, the same antibody clone in a different buffer formulation. Panel A : merged staining of anti-CD68 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, and counterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This image was generated using ab227688, the same clone, but with a different buffer formulation. Panel A : merged staining of anti-CD31 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, nuclear counterstain was DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD31 stained on endothelial cells and immune cell subsets with ab207090 at 1/500 dilution The section was incubated in three rounds of staining : in the order of ab207090 and ab16669 for 30 mins, then ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Panel A : merged staining of anti-CD19 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on human Hodgkin's lymphoma. Panel B : anti-CD19 staining the B lymphocytes in human Hodgkin's lymphoma. Panel C : anti-CD19 staining the B lymphocytes in human Hodgkin's lymphoma. Panel D : anti-CD3 staining the T lymphocytes in human Hodgkin's lymphoma. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab320735 at 1/2000 dilution, ab237772 at 1/5000 dilution and ab16669 at 1/150 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The secondary used was Opal Polymer HRP Ms + Rb. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This data was developed using ab227688, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human skeletal muscle labeling CD19 with ab227688 at 1/100 (0.15 ug/ml) dilution, followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control : no staining in human skeletal muscle.
The section was incubated with ab227688 for 10 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 rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Panel A : merged staining of anti-CD19 (green; Opal™520), anti-CD19 (green; Opal™570) and anti-CD3 (yellow; Opal™690) on human cerebrum. Panel B : anti-CD19 showed no staining in human cerebrum. Panel C : anti-CD19 showed no staining in human cerebrum. Panel D : anti-CD3 showed no staining in human cerebrum. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab320735 at 1/2000 dilution, ab237772 at 1/5000 dilution and ab16669 at 1/150 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The secondary used was Opal Polymer HRP Ms + Rb. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

Panel A : merged staining of anti-CD19 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on human tonsil. Panel B : anti-CD19 staining the B lymphocytes in human tonsil. Panel C : anti-CD19 staining the B lymphocytes in human tonsil. Panel D : anti-CD3 staining the T lymphocytes in human tonsil. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab320735 at 1/2000 dilution, ab237772 at 1/5000 dilution and ab16669 at 1/150 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The secondary used was Opal Polymer HRP Ms + Rb. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [SP291] - BSA and Azide free (AB237772)

This data was developed using ab227688, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD19 with ab227688 at 1/100 (0.15 ug/ml) dilution, followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in human tonsil.
The section was incubated with ab227688 for 10 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 rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.