Anti-CD21 antibody [SP186] - C-terminal

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD21 with ab227662 at 1/100 dilution (green) compared with a Rabbit IgG negative control (blue).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Flow Cytometry analysis of Raji (human Burkitt's lymphoma B lymphocyte) cells labeling CD21 with purified ab227662 at 1 : 20 dilution (5.3 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1 : 2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD21 with ab227662 at 1/100 dilution (1.06 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab227662 for 10 mins at room temperature.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Formalin-fixed, paraffin-embedded human colon tissue stained for CD21 using ab227662 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD21 using ab227662 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Formalin-fixed, paraffin-embedded human reactive lymph node tissue stained for CD21 using ab227662 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue sections labeling CD21 with ab227662 at 1/100 dilution (1.06 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human spleen, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab227662 for 10 mins at room temperature.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Formalin-fixed, paraffin-embedded human spleen tissue stained for CD21 using ab227662 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

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).

For mouse tissues, antigen retrieval was performed using high pH buffers (Dako, Agilent), followed by blocking of endogenous peroxidase activity with 3% hydrogen peroxide. Slides were then incubated with the primary antibody. Antigen/antibody complexes detection was carried out using a horseradish peroxidase-conjugated visualization system (Novolink Polymer, Leica). Staining was developed with 3,3′-diaminobenzidine (EnVision FLEX DAB Enhancer, Dako, Agilent), and nuclei were counterstained with hematoxylin (FLEX Hematoxylin, SM806, Dako, Agilent). Images were acquired using an Axiocam CCD camera (Zeiss) and processed with ZEN 2.1 software and Photoshop 9.0.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CD21 with ab227662 at 1/100 (0.28 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on mouse liver. The section was incubated with ab227662 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD21 with ab227662 at 1/100 (0.28 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse spleen. The section was incubated with ab227662 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling CD21 with ab227662 at 1/100 (0.28 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on mouse cardiac muscle. The section was incubated with ab227662 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue sections labeling CD21 with ab227662 at 1/100 dilution (1.06 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the rat spleen, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab227662 for 10 mins at room temperature.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD21 with ab227662 at 1/100 (0.28 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 μg/ml dilution (Green). membranous staining on mouse spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 μg/ml dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CD21 with ab227662 at 1/100 (0.28 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat spleen. The section was incubated with ab227662 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

Flow Cytometry analysis of 2.4G2 (rat B cell lymphoma B lymphocyte) cells labeling CD21 with purified ab227662 at 1 : 200 dilution (0.53 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1 : 2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

• WB

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Western blot - Anti-CD21 antibody [SP186] - C-terminal (AB227662)

All lanes:

Western blot - Anti-CD21 antibody [SP186] - C-terminal (ab227662) at 1/400 dilution

All lanes:

Daudi (Human Burkitt's lymphoma cell line) cell lysate

Predicted band size: 113 kDa

false