Anti-Carboxypeptidase A antibody [EPR24384-69]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Fluorescence multiplex immunohistochemical analysis of the human pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Carboxypeptidase A (ab278044, magenta; Opal™690), anti-Cytokeratin 19 (ab195872, green; Opal™520) and anti-Insulin (ab181547, red; Opal™570) on human pancreas. Panel B : anti-Carboxypeptidase A stained on acinar cells. Panel C : anti-Cytokeratin 19 stained on centroacinar cells and ducts. Panel D : anti-Insulin stained on beta cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab278044 at 1/4000 dilution (0.135 μg/ml), ab195872 at 1/8000 dilution (0.127 μg/ml), and ab181547 at 1/20000 dilution (0.053 μg/ml) 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. DAPI (blue) was used as a nuclear counter stain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Carboxypeptidase A with ab278044 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic staining on acinar cells of human pancreas (PMID : 23006325). The section was incubated with ab278044 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 solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Carboxypeptidase A with ab278044 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Negative control : No staining on human spleen. The section was incubated with ab278044 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 solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human pancreas labelling Somatostatin 28 with ab315106 at 1/7500 dilution (0.069 μg/ml) (B), GIP with ab271989 at 1/4000 dilution (0.27 μg/ml) (C) and Carboxypeptidase A with ab278044 at 1/4000 dilution (0.135 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain.

Panel A : merged staining of anti-Carboxypeptidase A (gray; Opal™690), anti-GIP (green; Opal™520) and anti-Somatostatin 28 (red; Opal™570) on human pancreas.

Panel B : anti-Somatostatin 28 stained on delta cells.

Panel C : anti-GIP stained on alpha cells.

Panel D : anti-Carboxypeptidase A stained on acinar cells.

The section was incubated in three rounds of staining : in the order of ab278044, ab271989, and ab315106 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Fluorescence multiplex immunohistochemical analysis of the human pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Carboxypeptidase A (ab278044, gray; Opal™690), anti-GIP (ab271989, green; Opal™520) and anti-Somatostatin 28 (ab111912, red; Opal™570) on human pancreas. Panel B : anti-Carboxypeptidase A stained on acinar cells. Panel C : anti-GIP stained on alpha cells. Panel D : anti-Somatostatin 28 stained on delta cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab278044 at 1/4000 dilution (0.135 μg/ml), ab271989 at 1/4000 dilution (0.27 μg/ml) and ab111912 at 1/9000 dilution (0.068 μg/ml) 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. DAPI (blue) was used as a nuclear counter stain.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Negative tissue image : IHC image of Carboxypeptidase A staining in a section of frozen normal human spleen performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab278044, 0.05 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

IHC image of Carboxypeptidase A staining in a section of frozen normal human pancreas* performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab278044, 0.05 ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining Carboxypeptidase A with ab278044 at a 1/4000 dilution, ab317319 anti-Claudin-3 used at 1/2000 dilution and ab314215 anti-C Peptide used at a 1/500 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-Carboxypeptidase A (green; Opal^™520), anti-Claudin 3 (magenta; Opal^™690) and anti-C Peptide (gray; Opal^™570) on rat pancreas. Panel B : anti-Carboxypeptidase A staining acinar cells in rat pancreas.
Panel C : ant-Claudin 3 staining cell membrane in rat pancreas.
Panel D : ant-C Peptide staining islet cells in rat pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab278044, ab317319 and ab314215 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse pancreas tissue labeling Carboxypeptidase A with ab278044 at 1/100 (5.39 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse pancreas is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 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-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Carboxypeptidase A with ab278044 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic staining on acinar cells of mouse pancreas. The section was incubated with ab278044 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 solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat pancreas tissue labeling Carboxypeptidase A with ab278044 at 1/500 (1.078 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) at 1/1000 dilution (Green). Positive staining on rat pancreas is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 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-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling Carboxypeptidase A with ab278044 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Cytoplasmic staining on acinar cells of rat pancreas. The section was incubated with ab278044 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 solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining C Peptide with ab314215 at a 1/500 dilution, ab278044 anti-Carboxypeptidase A used at 1/4000 dilution and ab52625 anti-Cytokeratin 19 used at a 1/6000 dilution.

Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-C Peptide (green; Opal^™520), anti-Carboxypeptidase A (magenta; Opal^™690) and anti-Cytokeratin 19 (grey; Opal^™570) on rat pancreas.
Panel B : anti-C Peptide staining beta cells in rat pancreas.
Panel C : anti-Carboxypeptidase A staining exocrine gland in rat pancreas.
Panel D : anti-Cytokeratin 19 staining pancreatic duct in rat pancreas.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab314215, ab278044 and ab52625 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution, ab278044 anti-Carboxypeptidase A used at 1/4000 dilution and ab181547 anti-Insulin used at a 1/20000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520), anti-Carboxypeptidase A (magenta; Opal™690) and anti-Insulin (gray; Opal™570) on mouse pancreas.

Panel B : anti-SLC7A5/LAT1 staining membrane in mouse pancreas.

Panel C : anti-Carboxypeptidase A staining acinar cells in mouse pancreas.

Panel D : anti-Insulin staining islet cells in mouse pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354, ab278044 and ab181547 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

Blocking and diluting buffer : 5% NFDM /TBST This antibody is specific to Carboxypeptidase A1 and has no cross reactivity with other family members tested Exposure time : 3.25 seconds

All lanes:

Western blot - Anti-Carboxypeptidase A antibody [EPR24384-69] (ab278044) at 1/1000 dilution

Lane 1:

Human Carboxypeptidase A1 recombinant protein (aa17-419) at 20 ng

Lane 2:

His tagged Human Carboxypeptidase A2 recombinant protein (aa20-419) at 20 ng

Lane 3:

His tagged Human Carboxypeptidase A4 recombinant protein (aa18-421) at 100 ng

Lane 4:

His tagged Human Carboxypeptidase A5 recombinant protein (aa36-416) at 200 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 3.25s

• WB

Lab

Western blot - Anti-Carboxypeptidase A antibody [EPR24384-69] (AB278044)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 21098023).

Activation of pro-carboxypeptidase A is initiated by proteolytic cleavage at the C-terminal end of the propeptide by trypsin and chymotrypsin (PMID : 8436102, 21098023).

Exposure time : 3 seconds.

All lanes:

Western blot - Anti-Carboxypeptidase A antibody [EPR24384-69] (ab278044) at 1/1000 dilution

Lane 1:

Human pancreas tissue lysate at 20 µg

Lane 2:

Mouse pancreas tissue lysate at 20 µg

Lane 3:

Rat pancreas tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 47 kDa

Observed band size: 33 kDa,43 kDa

false