Anti-CD19 antibody [EPR23174-145]

31 product images

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  Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Flow cytometric analysis of mouse peripheral blood mononuclear cell (PBMC) cells labeling CD19 with ab245235 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
  

  Cells were stained with rabbit IgG (Left) or ab245235 (Right). Then stained with anti-CD3 conjugated to Alexa Fluor^® 647. CD19 and CD3 are mutually exclusive expressed in mouse PBMC.

  Gated on viable cells.

• 

  Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized perfusion fixed frozen Mouse spleen tissue labeling CD19 with ab245235 at 1/250 (1.82 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).
  

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

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

• 

  Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Blocking and dilution buffer: 5% NFDM/TBS.
  

  Exposure time: 48 seconds.

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).

  Negative control: CTLL-2 (PMID: 1370948) is treated as a negative cell line according to the literature, that CD19 is a B lymphocyte cell-surface marker that is expressed early during pre-B-cell differentiation, not in T cells.

  All lanes: Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution

  Lane 1: WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate at 20 µg

  Lane 2: A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate at 20 µg

  Lane 3: CTLL-2 (mouse T lymphocyte), whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 61 kDa

  Observed band size: 60-120 kDa

• 

  Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)

  CD19 was immunoprecipitated from 0.35 mg mouse spleen lysate with ab245235 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245235 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Mouse spleen lysate 10μg

  Lane 2: ab245235 IP in mouse spleen lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab245235 in mouse spleen lysate

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

  Exposure time: 30 seconds.

  All lanes: Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Predicted band size: 61 kDa

  Observed band size: 60-120 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD19 with ab245235 at 1/1000 dilution (0.446 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on mouse spleen (PMID: 18648532). The section was incubated with ab245235 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labeling CD19 with ab245235 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in some subsets of mouse PBMCs. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• 

  Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Exposure time: 48 seconds.

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).

  All lanes: Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution

  All lanes: Mouse spleen tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 61 kDa

  Observed band size: 60-120 kDa

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining MARCO with Anti-MARCO antibody [RM2074] ab322705 at a 1:500 (1.032 ug/ml) dilution, Anti-F4/80 antibody [EPR26545-166] ab300421 anti-F4/80 used at 1:5000 (0.10 ug/ml) dilution and ab245235 anti-CD19 used at a 1:1000 (0.444 ug/ml) dilution.

  Panel A: merged staining of anti-MARCO (green; Opal™520), anti-F4/80 (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on mouse liver.
  
  Panel B: anti-MARCO staining Kupffer cells in mouse liver.
  
  Panel C: anti-F4/80 staining Kupffer cells in mouse liver.
  
  Panel D: anti-CD19 staining B lymphocytes in mouse liver.
  
  Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-MARCO antibody [RM2074] ab322705, Anti-F4/80 antibody [EPR26545-166] ab300421 and ab245235 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.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse bone marrow staining of F4/80 with Anti-F4/80 antibody [EPR26545-166] ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse bone marrow.
  Panel B: anti-F4/80 staining macrophages in mouse bone marrow.
  Panel C: anti-CD19 staining B lymphocytes in mouse bone marrow.
  Panel D: anti-CD3 staining T lymphocytes in mouse bone marrow.
  Nuclear DNA was labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-F4/80 antibody [EPR26545-166] ab300421, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus staining of NCR1 with Anti-NCR1 antibody [EPR23097-35] ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse thymus.
  Panel B: anti-NCR1 staining natural killer cells 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-NCR1 antibody [EPR23097-35] ab233558, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node staining of NCR1 with Anti-NCR1 antibody [EPR23097-35] ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse lymph node.
  Panel B: anti-NCR1 staining natural killer cells 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-NCR1 antibody [EPR23097-35] ab233558, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen staining of NCR1 with Anti-NCR1 antibody [EPR23097-35] ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse spleen.
  Panel B: Anti-NCR1 staining natural killer cells 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-NCR1 antibody [EPR23097-35] ab233558, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen staining of F4/80 with Anti-F4/80 antibody [EPR26545-166] ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse spleen.
  Panel B: anti-F4/80 staining macrophages 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-F4/80 antibody [EPR26545-166] ab300421, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node staining of F4/80 with Anti-F4/80 antibody [EPR26545-166] ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse lymph node.
  Panel B: anti-F4/80 staining macrophages 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-F4/80 antibody [EPR26545-166] ab300421, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus staining of F4/80 with Anti-F4/80 antibody [EPR26545-166] ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

  Panel A: merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse thymus.
  Panel B: anti-F4/80 staining macrophages 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 labelled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-F4/80 antibody [EPR26545-166] ab300421, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue staining Neutrophil Elastase with Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

  Panel A: merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B: anti-Neutrophil Elastase staining neutrophils 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 Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus tissue staining Neutrophil Elastase with Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

  Panel A: merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B: anti-Neutrophil Elastase staining neutrophils 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 Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse bone marrow tissue staining Neutrophil Elastase with Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

  Panel A: merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B: anti-Neutrophil Elastase staining neutrophils 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 Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node tissue staining Neutrophil Elastase with Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with Anti-CD3 epsilon antibody [CAL57] ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

  Panel A: merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B: anti-Neutrophil Elastase staining neutrophils 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 Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.

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

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  CD19 immunohistochemistry staining of mouse spleen using rabbit anti-CD19 antibody
  
  

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD21+CD35 with Anti-CD21+CD35 antibody [RM2062] ab318999 at a 1:2000 (0.255 ug/ml) dilution, ab245235 anti-CD19 used at 1:1000 (0.444 ug/ml) dilution and Anti-CD3 epsilon antibody [SP7] ab16669 anti-CD3 used at a 1:150 (0.06 ug/ml) dilution.

  Panel A: merged staining of anti-CD21+CD35 (green; Opal™520), anti-CD19 (magenta; Opal™690) and anti-CD3 (yellow; Opal™570) on mouse spleen.
  
  Panel B: anti-CD21+CD35 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 Anti-CD21+CD35 antibody [RM2062] ab318999, ab245235 and Anti-CD3 epsilon antibody [SP7] ab16669 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.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  CD19 immunohistochemistry staining of mouse spleen using rabbit anti-CD19 antibody
  
  

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD272/BTLA with Anti-CD272/BTLA antibody [EPR29068-115] ab317827 at a 1:1000 (0.44 ug/ml) dilution, Anti-CD272/BTLA antibody [EPR29068-115] ab317827 anti-BTLA used at 1:100 (4.92 ug/ml) dilution and Anti-CD3 epsilon antibody [SP7] ab16669 anti-CD3 used at a 1:150 (0.06 ug/ml) dilution.

  Panel A: merged staining of anti-CD19 (magenta; Opal™690), anti-BTLA (green; Opal™520) and anti-CD3 (yellow; Opal™570) on mouse spleen.
  
  Panel B: anti-CD19 staining B lymphocytes in mouse spleen.
  
  Panel C: anti-BTLA staining B lymphocytes and T 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 ab245235, Anti-CD272/BTLA antibody [EPR29068-115] ab317827 and Anti-CD3 epsilon antibody [SP7] ab16669 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.

• 

  This image is courtesy of an anonymous customer review

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed mouse spleen tissue. Stained with ab245235 at 1/1000 dilution. Secondary antibody used was Dako EnVision+ System- HRP Labelled Polymer Anti-R. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0.

• 

  This image is courtesy of an anonymous customer review

  Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Immunohistochemistry (Frozen sections) analysis of acetone-fixed mouse spleen tissue staining with ab245235 at 1/1000 dilution. Secondary antibody was Dako EnVision+ System- HRP Labelled Polymer Anti-R Samples were incubated with the primary antibody with 5% Normal Goat Serum for 19 hours at 4°C. Blocking was done using 5% serum for 1 hour at 21°C

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining LY6C with Anti-LY6C antibody [RM1151] ab317272 at a 1:100 (0.978 ug/ml) dilution, Anti-CD11b antibody [EPR1344] ab133357 anti-CD11b used at 1:20000 (0.05 ug/ml) dilution and ab245235 anti-CD19 used at a 1:1000 (0.999 ug/ml) dilution.

  Panel A: merged staining of anti-Ly6c (green; Opal™690), anti-CD11b (magenta; Opal™570) and anti-CD19 (yellow; Opal™520) on mouse spleen.
  
  Panel B: anti-Ly6c stained on monocytes/macrophages.
  
  Panel C: anti-CD11b stained on monocytes/macrophages.
  
  Panel D: anti-CD19 stained on B cells.
  
  Co-staining of Ly6c and CD11b can be observed.
  
  The section was incubated in three rounds of staining: in the order of Anti-LY6C antibody [RM1151] ab317272, Anti-CD11b antibody [EPR1344] ab133357, and ab245235 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
  
  Nuclear DNA was labeled with DAPI (shown in blue).

  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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen treated with lipopolysaccharide (1 ug/ml) for 16 hours and brefeldin a (1 ug/ml) for overnight tissue staining CD80 with ab322922 at a 1:2000 (0.257 ug/ml) dilution, ab245235 anti-CD19 used at 1:1000 (0.444 ug/ml) dilution.

  Panel A: merged staining of anti-CD80 (green; Opal™520) and anti-CD19 (magenta; Opal™570) on mouse spleen treated with Lipopolysaccharide.
  
  Panel B: anti-CD80 staining immune cells in mouse spleen treated with Lipopolysaccharide.
  
  Panel C: ant-CD19 staining B lymphocytes in mouse spleen treated with Lipopolysaccharide.
  
  Panel D: Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of ab322922 and ab245235 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.

• 

  Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Flow cytometry overlay histogram showing CD3+ve mouse splenocytes stained with ab245235 (right) and with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control, used at the same concentration and conditions as the primary antibody (left). The cells were incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab245235) (1x 10⁶ in 100μl at 1.0 μg/ml (1/500)) for 30min on ice.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 for 30min on ice

  Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD23 with Anti-CD23 antibody [EPR28712-26] ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with Anti-CD3 epsilon antibody [CAL57] 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 Anti-CD23 antibody [EPR28712-26] ab315289, ab245235 and Anti-CD3 epsilon antibody [CAL57] 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.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lymph node tissue staining CD23 with Anti-CD23 antibody [EPR28712-26] ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with Anti-CD3 epsilon antibody [CAL57] 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 Anti-CD23 antibody [EPR28712-26] ab315289, ab245235 and Anti-CD3 epsilon antibody [CAL57] 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.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thymus tissue staining CD23 with Anti-CD23 antibody [EPR28712-26] ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with Anti-CD3 epsilon antibody [CAL57] 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 Anti-CD23 antibody [EPR28712-26] ab315289, ab245235 and Anti-CD3 epsilon antibody [CAL57] 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.

• 

  Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Flow cytometry staining of C57BL/6 mouse splenocytes with ab245235 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10µg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab245235 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶in 100 µl at 1.0 µg/ml (1/2120)) for 30min on ice. The cells were simultaneously stained with CD3.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

  Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• 

  Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining MARCO with Anti-MARCO antibody [RM2074] ab322705 at a 1:500 (1.032 ug/ml) dilution, ab245235 anti-CD19 used at 1:1000 (0.444 ug/ml) dilution and Anti-CD3 epsilon antibody [CAL57] ab237721 anti-CD3 used at a 1:2000 (0.264 ug/ml) dilution.

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

  The section was incubated in three rounds of staining: in the order of Anti-MARCO antibody [RM2074] ab322705, ab245235 and Anti-CD3 epsilon antibody [CAL57] ab237721 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.