Anti-PSGL-1 antibody [EPR28170-27]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse glioblastoma. The section was incubated with ab316113 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

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Flow Cytometry - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Flow cytometric analysis of Mouse splenocytes cells labelling PSGL-1 with ab316113 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Gated on viable cells. Cells are co-stained with CD3 conjugated to Alexa Fluor®647.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab316113 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-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of rat colon. The section was incubated with ab316113 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-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse colon. The section was incubated with ab316113 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

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Immunocytochemistry/ Immunofluorescence - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse splenocyte cells labelling PSGL-1 with ab316113 at 1/50 (10.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing membrane staining in a subset of mouse splenocytes.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Anti-rat CD3 mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain at 1/100 (5ug/ml) dilution (Red). 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 (2 ug/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab316113 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.

• WB

Supplier Data

Western blot - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

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

Low expression : skeletal muscle, kidney (PMID 8639776).

The molecular weight observed is consistent with what has been described in the literature (PMID : 18519846, 9660879).

PSGL-1(P-selectin is the P-selectin glycoprotein ligand-1) is a 125KD glycoprotein and the 250KD band represents the dimer.

The identity of the bands below 75 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-PSGL-1 antibody [EPR28170-27] (ab316113) at 1/1000 dilution

Lane 1:

Rat spleen tissue lysate at 20 µg

Lane 2:

Rat lymph node tissue lysate at 20 µg

Lane 3:

Rat skeletal muscle tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 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

Observed band size: 125 kDa,250 kDa,124 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

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

Low expression : EL4.IL-2 (PMID : 34090013), NIH/3T3; skeletal muscle (PMID 8639776), kidney (PMID 8639776).

The molecular weight observed is consistent with what has been described in the literature (PMID : 18519846, 9660879).

PSGL-1(P-selectin is the P-selectin glycoprotein ligand-1) is a 125KD glycoprotein and the 250KD band represents the dimer.

The identity of the bands below 75 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lane 1-4 : 81 seconds; Lane 5-9 : 10 seconds

All lanes:

Western blot - Anti-PSGL-1 antibody [EPR28170-27] (ab316113) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

A20 (mouse reticum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse lymph node tissue lysate at 20 µg

Lane 7:

Mouse thymus tissue lysate at 20 µg

Lane 8:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 9:

Mouse kidney tissue lysate at 20 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

Observed band size: 125 kDa,250 kDa,124 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on mouse skeletal muscle (PMID : 8639776). The section was incubated with ab316113 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-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse PSGL-1 expression vector containing a Myc-His tag. (B) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a Myc-His-tagged mouse PSGL-1 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab316113 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-PSGL-1 antibody [EPR28170-27] (AB316113)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling PSGL-1 with ab316113 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on rat skeletal muscle. The section was incubated with ab316113 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.