Anti-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Peptide YY/PYY with ab324525 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on enteroendocrine cells in human colon (PMID : 25724002).
The primary antibody was incubated 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-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a PYY expression vector containing a His tag cell pellet. (B) HEK-293T transfected with a NPY expression vector containing a His tag cell pellet. (C) HEK-293T transfected with an empty vector containing a His tag cell pellet. tissue labeling Peptide YY/PYY with ab324525 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a PYY expression vector containing a His tag cell pellet, no staining on (B) HEK-293T transfected with a NPY expression vector containing a His tag cell pellet and (C) HEK-293T transfected with an empty vector containing a His tag cell pellet.
The primary antibody was incubated 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-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HuTu 80 (human Small intestine; Duodenum epithelial) cells labelling Peptide YY/PYY with ab324525 at 1/50 (10.16 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing increased cytoplasmic staining in HuTu 80 cells (shown in green) treated with sodium butyrate (5 mM) for 48 hr. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324525 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HuTu 80 (human Small intestine; Duodenum epithelial) treated with 5mM Sodium butyrate for 48 hours (Green) / Untreated control (Magenta) cells labelling Peptide YY/PYY with ab324525 at 1/500 dilution (0.1ug) (Magenta) and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining GC-C with ab323804 at a 1/100 dilution, ab324525 anti-Peptide YY used at 1/2000 dilution and ab272692 anti-MUC2 used at a 1/2000 dilution.

Panel A : anti-GC-C (green; Opal™520), anti-Peptide YY (magenta; Opal™690) and anti-MUC2 (gray; Opal™570) on human colon.

Panel B : anti-GC-C staining apical membrane of epithelium in human colon.

Panel C : anti-Peptide YY staining enteroendocrine cells in human colon.

Panel D : anti-MUC2 staining goblet cells in human colon.

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

The section was incubated in three rounds of staining : in the order of ab323804, ab324525 and ab272692 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

"

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Peptide YY/PYY with ab324525 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human liver.
The primary antibody was incubated 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-Peptide YY/PYY antibody [EPR30490-578] - BSA and Azide free (AB324534)

"

This data was developed using ab324525, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Peptide YY/PYY with ab324525 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human spleen.
The primary antibody was incubated 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

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