Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free

• Flow Cyt

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Flow Cytometry - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) cell line labeling PSGL-1 using ab227836 at 1/500 dilution (Right). Cells were stained with rabbit monoclonal IgG (ab172730) (Left) or ab227836 (Right). Then stained with anti-CD3 conjugated to Alexa Fluor^® 647. Gated on viable cells. The secondary antibody was a Goat anti rabbit IgG (Alexa Fluor^® 488, ab150097) at 1/2000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PSGL-1 with ab227836 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on human tonsil (PMID : 19594630, 16567389). The section was incubated with ab227836 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells stained for PSGL-1 using ab227836 (Green) at 1/50 dilution, followed by an AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing membranous staining in Jurkat cells. The nuclear counterstain was DAPI (Blue). Tubulin was stained using the Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling PSGL-1 with ab227836 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on microglial cells of human cerebrum is observed. The section was incubated with ab227836 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling PSGL-1 with ab227836 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on infiltrated leukocytes of human colon (PMID : 19594630). The section was incubated with ab227836 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-PSGL-1 antibody [EPR22504-36] - BSA and Azide free (AB255290)

PSGL-1 was immunoprecipitated form 0.35 mg of Jurkat (human T cell leukemia T lymphocyte) whole cell lysate using ab227836 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab227836 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary at a 1/1000 dilution.

Lane 1 : Jurkat whole cell lysate μg (input).
Lane 2 : ab227836 IP in jurkat whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227836 in Jurkat whole cell lysate.

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

Exposure time : 5 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227836).

The expression profile observed is consistent with what has been described in the literature (PMID : 9660879; PMID : 28515724). PSGL-1(P-selectin is the P-selectin glycoprotein ligand-1) is a 125KD glycoprtein and the 250KD band represents the dimer.

All lanes:

Immunoprecipitation - Anti-PSGL-1 antibody [EPR22504-36] (<a href='/en-us/products/primary-antibodies/psgl-1-antibody-epr22504-36-ab227836'>ab227836</a>)

Predicted band size: 43 kDa

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