Anti-CD20 antibody [SP32] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

This data was developed using ab64088, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD20 with ab64088 at 1/100 dilution.

Positive staining on human tonsil.

The section was incubated with ab64088 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Sodium-Citrate buffer (pH 6.0, Epitope Retrieval Solution1) for 10 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Flow cytometry analysis of Ramos (human Burkitt's lymphoma ) labeling CD20 with purified ab64088 at 1/40 dilution (7.78 μg/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 (ab64088).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Flow cytometric analysis of rabbit anti-CD20 (SP32) antibody ab64088(1/100)  in Jurkat cells (green) compared to negative control of rabbit IgG (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64088).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) cells labeling CD20 with purified ab64088 at 1/100 (3.1 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64088).

• IHC-P

AbReview83784****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Immunohistochemistry analysis of paraformaldehyde fixed Human tonsil tissue staining CD20 with ab236434 at 1/400 and Alexa Fluor 555 donkey anti rabbit secondary. The tissue was incubated with the primary antibody for 16 hours at 4 degrees with PBS + 2% FCS. 5% normal human serum was used for blocking for 1 hour at room temperature.

This image is courtesy of an Abreview submitted by Natalie Papazian

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

This data was developed using ab64088, 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).

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.

• IP

Lab

Immunoprecipitation - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

ab64088 at 1/30 dilution immunoprecipitating CD20 in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate. Western blot was performed from the immunoprecipitate using ab64088 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution. Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 µg Lane 2 : ab64088 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab64088 in Raji whole cell lysate Blocking/Dilution buffer : 5% NFDM/TBST. Exposure time : 8 seconds. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab64088).

Lane 2:

Immunoprecipitation - Anti-CD20 antibody [SP32] (<a href='/en-us/products/primary-antibodies/cd20-antibody-sp32-ab64088'>ab64088</a>) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-CD20 antibody [SP32] - BSA and Azide free (ab236434)

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 25 kDa,33 kDa

false

Exposure time: 8s

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue labeling Sialoadhesin/CD169, CD3 epsilon and CD20 with ab312840 at 1/100 dilution, ab16669 at 1 : 150 dilution and ab236434 at 1 : 5000 dilution. Panel A : merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD20 (gray; Opal™570) on mouse spleen. Panel B : anti-Sialoadhesin/CD169 stained on macrophages. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD20 stained on B cells. The section was incubated in three rounds of staining : in the order of ab312840, ab16669, and ab236434 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

This data was developed using ab64088, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CD20 with ab64088 at 1/100 dilution.

Positive staining on rat spleen.

The section was incubated with ab64088 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Sodium-Citrate buffer (pH 6.0, Epitope Retrieval Solution1) for 10 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

This data was developed using ab64088, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD20 with ab64088 at 1/100 dilution.

Positive staining on mouse spleen.

The section was incubated with ab64088 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Sodium-Citrate buffer (pH 6.0, Epitope Retrieval Solution1) for 10 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat spleen tissue labeling Sialoadhesin/CD169, CD3 epsilon and CD20 with ab312840 at 1/100 dilution, ab16669 at 1 : 150 dilution and ab236434 at 1 : 5000 dilution. Panel A : merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD20 (gray; Opal™570) on rat spleen. Panel B : anti-Sialoadhesin/CD169 stained on macrophages. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD20 stained on B cells. The section was incubated in three rounds of staining : in the order of ab312840, ab16669, and ab236434 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-CD20 antibody [SP32] - BSA and Azide free (AB236434)

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CD20 antibody [SP32] - BSA and Azide free (AB236434)

Flow cytometry analysis of WEHI-231 (mouse lymphoblast B cell lymphoma) labeling CD20 with purified ab64088 at 1/40 dilution (7.78 μg/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 (ab64088).