Anti-PAR2 antibody [EPR13675] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180953).

Flow cytometry overlay histogram showing left HT-29 positive cells and right negative Daudi stained with ab180953 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab180953) (1x 106 in 100μl at 5.0μg/ml (1/560)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling PAR2 with Purified ab180953 at 1/50 dilution (2.76 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling PAR2 with unpurified ab180953 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling PAR2 with Purified ab180953 at 1/100 dilution (1.38 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling PAR2 with Purified ab180953 at 1/100 dilution (1.38 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling PAR2 with Purified ab180953 at 1/100 dilution (1.38 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Unknown

Western blot - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.

We are unsure how to define the extra bands.

All lanes:

Western blot - Anti-PAR2 antibody [EPR13675] (<a href='/en-us/products/primary-antibodies/par2-antibody-epr13675-ab180953'>ab180953</a>) at 1/1000 dilution

Lane 1:

Human kidney lysate at 15 µg

Lane 2:

Mouse kidney lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 44 kDa

false

• WB

Unknown

Western blot - Anti-PAR2 antibody [EPR13675] - BSA and Azide free (AB240232)

This data was developed using ab180953, the same antibody clone in a different buffer formulation.

We are unsure how to define the extra bands.

All lanes:

Western blot - Anti-PAR2 antibody [EPR13675] (<a href='/en-us/products/primary-antibodies/par2-antibody-epr13675-ab180953'>ab180953</a>) at 1/1000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Rat kidney lysate at 15 µg

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: 44 kDa

false