Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling PGRPS with ab325131 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

Cells were stained with anti-CD3 conjugated to Alexa Fluor®647. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse lung (PMID : 9707603). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen (PMID : 9707603). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cerebrum (PMID : 9707603). 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse bone marrow cells labelling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in mouse CD11B+ bone marrow cells (shown in green). 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).

ab8878 Anti-CD11b Rat monoclonal antibody was used to counterstain tubulin at 1/100 dilution (Magenta). 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 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse blood cells labelling PGRPS with ab325131 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

Cells were stained with anti-CD3 conjugated to Alexa Fluor®647. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cardiac muscle (PMID : 9707603). 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse bone marrow tissue labeling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse bone marrow. 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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling PGRPS with ab325131 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

Cells were stained with anti-Gr-1 conjugated to Pacific Blue. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse blood cells labelling PGRPS with ab325131 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

Cells were stained with anti-Gr-1 conjugated to Pacific Blue. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• WB

Lab

Western blot - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

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

Low expression : heart (PMID : 9707603 ; PMID : 11461926; PMID : 9707603 ), NIH/3T3

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9707603 ; PMID : 11461926; PMID : 9707603 ).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-PGRPS antibody [EPR30470-91] (<a href='/ko/products/primary-antibodies/pgrps-antibody-epr30470-91-ab325131'>ab325131</a>) at 1/1000 dilution

Lane 1:

Mouse bone marrow tissue lysate at 48 µg

Lane 2:

Mouse spleen tissue lysate at 48 µg

Lane 3:

Mouse heart tissue lysate at 48 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 48 µg

Secondary

All lanes:

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

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling PGRPS with ab325131 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black 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.

Negative control : NIH/3T3

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGRPS antibody [EPR30470-91] – BSA and Azide free (AB325145)

This data was developed using ab325131, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling PGRPS with ab325131 at 1/1000 (0.522 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Negative control : confocal image showing no staining in NIH/3T3 cell line (shown in green). 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).

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