Anti-CD23 antibody [EPR28712-26] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using the same antibody clone in a different buffer formulation (ab315289).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse spleen.
Panel B : anti-CD23 staining B lymphocytes in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling CD23 with ab315289 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cardiac muscle. The section was incubated with ab315289 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 was performed 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-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD23 with ab315289 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on mouse spleen. The section was incubated with ab315289 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 was performed 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-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tumor tissue labeling CD23 with ab315289 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on several immune cells of mouse pancreas tumor. The section was incubated with ab315289 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse splenocytes cells labelling CD23 with ab315289 at 1/100 (5.06 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing membranous staining in subsets of mouse splenocytes. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse splenocytes cells labelling CD23 with ab315289 at 1/500 dilution (0.1 ug, Right) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with CD19 conjugated to Alexa Fluor®647. Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse splenocytes cells labelling CD23 with ab315289 at 1/500 dilution (0.1 ug, Right) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with CD3 conjugated to Alexa Fluor®647. Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CD23 with ab315289 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse skeletal muscle. The section was incubated with ab315289 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using the same antibody clone in a different buffer formulation (ab315289).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lymph node tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse lymph node.
Panel B : anti-CD23 staining B lymphocytes in mouse lymph node.
Panel C : anti-CD19 staining B lymphocytes in mouse lymph node.
Panel D : anti-CD3 staining T lymphocytes in mouse lymph node.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using the same antibody clone in a different buffer formulation (ab315289).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thymus tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse thymus.
Panel B : anti-CD23 staining B lymphocytes in mouse thymus.
Panel C : anti-CD19 staining B lymphocytes in mouse thymus.
Panel D : anti-CD3 staining T lymphocytes in mouse thymus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 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.

• IP

Supplier Data

Immunoprecipitation - Anti-CD23 antibody [EPR28712-26] - BSA and Azide free (AB315290)

This data was developed using ab315289, the same antibody clone in a different buffer formulation.

CD23 was immunoprecipitated from 0.35 mg Mouse spleen whole cell lysate with ab315289 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315289 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse spleen whole cell lysate

Lane 2 : ab315289 IP in Mouse spleen whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315289 in mouse spleen whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-CD23 antibody [EPR28712-26] (<a href='/en-us/products/primary-antibodies/cd23-antibody-epr28712-26-ab315289'>ab315289</a>) at 1/30 dilution

All lanes:

Mouse spleen whole cell lysate

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

false

Exposure time: 125s